the biotechnology education company ® - austin community college
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The Biotechnology Education Company ®
EDVOTEK, Inc. • 1-800-EDVOTEK • www.edvotek.com
EVT 001197AM
EDVO-Kit #
102& 102-Q
Restriction Enzyme Cleavage of DNA
Storage: See Page 3 for specific storage instructions
ExpERImENT OBjECTIVE:
The objective of this experiment is to develop anunderstanding of the use of restriction endonucleases
as tools to cut DNA at specific sequences.
This document includes instructions for both EDVOTEK Experiment # 102 and # 102-Q. Please follow instructions for the appropriate experiment. Experiment #102 is designed for DNA staining with InstaStain® Methylene Blue. Experiment # 102-Q is designed for DNA staining with InstaStain® Ethidium Bromide.
Restriction Enzyme Cleavage of DNA
EVT 001197AM
2
102102EDVO-Kit #
EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
All components are intended for educational research only. They are not to be used for diagnostic or drug purposes, nor adminis-tered to or consumed by humans or animals.
THIS EXPERIMENT DOES NOT CONTAIN HUMAN DNA. None of the experiment components are derived from human sources.
EDVOTEK, The Biotechnology Education Company, and InstaStain are registered trademarks of EDVOTEK, Inc.. Ready-to-Load and UltraSpec-Agarose are trademarks of EDVOTEK, Inc.
Page
Experiment Components 3Experiment Requirements 3Background Information 5
Experiment procedures Experiment Overview and General Instructions 9
Agarose Gel Electrophoresis Agarose Gel Requirements for this Experiment 12 Preparing the Gel Bed 12 Casting Agarose Gels 12 Preparing the Gel for Electrophoresis 13 Loading the Samples 14
Running the Gel 15 Staining and visualization of DNA 15 Study Questions 16 Instructor's Guidelines 17 Notes to the Instructor 18 Pre-Lab Preparations 21 Experiment Results and Analysis 23 Study Questions and Answers 24
Appendices 0.8% Agarose Gel Preparation A DNA Staining with InstaStain® Methylene Blue 26 B DNA Staining with InstaStain® Ethidium Bromide 27
C Quantity Preparations for Agarose Gel Electrophoresis 28 Staining and Visualization of DNA D Method 1: InstaStain® MetBlue One-step Staining and destaining 29 E Method 2: InstaStain® MetBlue Cards 30 F Method 3: Liquid Methylene Blue Plus™ 32 G Method 4: InstaStain® Ethidium Bromide Cards 33
Material Safety Data Sheets 34
This document includes instructions for both EDVOTEK Experiment # 102 and # 102-Q. Please follow in-structions for the appropriate experiment. Experiment # 102 is designed for DNA staining with InstaStain® Methylene Blue. Experiment # 102-Q is designed for DNA staining with InstaStain® Ethidium Bromide.
Table of Contents
Restriction Enzyme Cleavage of DNA
EVT 001197AM
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FAx: (�01) �40-0582 • email: [email protected]
102EDVO-Kit #
Experiment Components
Requirements
READy-TO-LOAD™ DNA sAmpLEs FOR ELECTROphOREsIs
A Plasmid DNA (uncut) B Plasmid cut with Bgl l C Plasmid cut with Eco Rl D Lambda DNA (uncut) E Lambda DNA cut with Eco Rl F Lambda DNA cut with Bgl l
REAGENTs & suppLIEs
• UltraSpec-Agarose™ powder • Concentrated electrophoresis buffer • Practice Gel Loading Solution • 1 ml pipet • 100 ml graduated cylinder (packaging for samples) • Microtipped Transfer Pipets • DNA Stain for Standard Series 100 experiments • InstaStain® Methylene Blue • Methylene Blue Plus™ • DNA Stain for Series 100-Q experiments • InstaStain® Ethidium Bromide
• Horizontal gel electrophoresis apparatus • D.C. power supply • Automatic micropipets with tips • Balance • Microwave, hot plate or burner • Pipet pump • 250 ml flasks or beakers • Hot gloves • Safety goggles and disposable laboratory gloves • Distilled or deionized water • For gel staining with InstaStain® Methylene Blue • Small plastic trays or large weigh boats for destaining • White light DNA visualization system • For gel staining with InstaStain® Ethidium Bromide • UV Transilluminator • Photodocumentation system (optional)
Store entire experiment at
room temperature.
DNA samples do not require heating prior to gel loading.
DNA samples are stable at room temperature. However, if the experiment will not be conducted within one month of receipt, it is recommended that the DNA samples be stored in the refrigerator.
Over time, some evaporation of samples may occur. Before distributing reagents to stu-dents, check sample volumes as described in the Instructor’s Pre-Lab Preparation section.
Components & Requirements
Restriction Enzyme Cleavage of DNA
EVT 001197AM
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102102EDVO-Kit #
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Please have the following information ready:
• Experiment number and title• Kit lot number on box or tube• Literature version number (in lower right corner)• Approximate purchase date
Technical ServiceDepartment
This document includes instructions for both EDVOTEK Experiment # 102 and # 102-Q. Please follow instructions for the appropriate experiment.
Experiment #102
Designed for DNA staining and visualization with InstaStain® Methylene Blue. This experiment requires a 0.8% gel with the following volume:
30 ml (7 x 7 cm) or 60 ml (7 x 14 cm)
Refer to Table A.1 or A.2 in Appendix A for agarose gel preparation specifi-cations.
Experiment # 102-Q
Designed for DNA staining and visualization with InstaStain® Ethidium Bro-mide. This experiment requires a 0.8% gel with the following volume:
25 ml (7 x 7 cm) or 50 ml (7 x 14 cm)
Refer to Table A.3 or A.4 in Appendix B for agarose gel preparation specifi-cations.
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Restriction Enzyme Cleavage of DNA
The discovery of restriction enzymes ushered in a new era of molecular genetics. These enzymes cut the DNA molecule in a highly specific and reproducible way. This, in turn, has lead to the development of molecular cloning and the mapping of genetic structures.
Restriction enzymes are endonucleases which catalyze the cleavage of the phosphodiester bonds within both strands of DNA. They require Mg+2 for activity and generate a 5 prime (5') phosphate and a 3 prime (3') hydroxyl group at the point of cleavage. The distinguishing feature of restriction en-zymes is that they only cut at very specific sequences of bases. Restriction enzymes are obtained from many different species of bacteria (including blue-green algae). To date, over 3,000 restriction enzymes have been discovered and catalogued.
Restriction enzymes are named according to the organism from which they are isolated. This is done by using the first letter of the genus followed by the first two letters of the species. Only certain strains or sub-strains of a particular species may produce restriction enzymes. The type of strain or substrain sometimes follows the species designation in the name. Finally, a Roman numeral is always used to designate one out of possibly several different restriction enzymes produced by the same organism or by different substrains of the same strain.
A restriction enzyme requires a specific double stranded recognition se-quence of nucleotides to cut DNA. Recognition sites are usually 4 to 8 base pairs in length. Cleavage occurs within or near the site. The cleavage posi-tions are indicated by arrows. Recognition sites are frequently symmetrical, i.e., both DNA strands in the site have the same base sequence when read 5' to 3'. Such sequences are called palindromes. Consider the recognition site and cleavage pattern of Eco RI as an example.
↓ 5'-GAATTC-3' 5'-G AATTC-3' 3'-CTTAAG-5' 3-CTTAA G-5' ↑
As shown above, Eco RI causes staggered cleavage of its site. The ends of the DNA fragments are called “sticky” or “cohesive” ends because the single-stranded regions of the ends are complementary.
Restriction Enzyme Organism
Bgl l Bacillus globigii Bam HI Bacillus amyloliquefaciens H
Eco Rl Escherichia coli, strain RY 13
Eco Rll Escherichia coli, strain R 245
Hae III Haemophilus aegyptius
Hind III Haemophilus influenzae Rd
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/labora-tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1989,1992,1994,1997,1998, 2000, 2004, 2007, EDVOTEK, Inc., all rights reserved. EVT 001197AM
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102Restriction Enzyme Cleavage of DNA
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Restriction Enzyme Cleavage of DNA
Some restriction enzymes, such as Hae III, introduce cuts that are opposite each other. This type of cleavage generates “blunt” ends.
↓ 5'-GGCC-3' 5'-GG CC-3' 3'-CCGG-5' 3'-CC GG-5' ↑
The recognition sites of some restriction enzymes contain variable base posi-tions. For example, Ava I recognizes:
↓ 5'-CPyCGPuG-3' (Py = pyrimidine = C or T and 3'-GPuGCPyC-5' Pu = purine = G or A) ↑
Keep in mind that A pairs with T and G pairs with C. Consequently, there are four possible sequences Ava I recognizes. Recognition sites of this type are called degenerate.
There are some recognition sites that are divided by a certain number of totally variable bases. For example, Bgl I recognizes:
↓ 5'-GCCNNNNNGGC-3' (N = A, G, C or T) 3'-CGGNNNNNCCG-5' ↑
There are 625 possible sequences Bgl I can cleave. The only bases the enzyme truly “recognizes” are the six G-C base pairs at the ends, which forms a palindrome. In the case of Bgl I, these true recognition bases must always be separated by 5 base pairs of DNA, otherwise the enzyme cannot properly interact with the DNA and cleave it. Rec-ognition sites like that of Bgl I are called hyphen-ated sites.
In general, the longer the DNA molecule, the greater the probability that a given recognition site will occur. Therefore, human chromosomal DNA, which contains three billion base pairs, has many more recognition sites than a plasmid DNA containing only several thousand base pairs. How-ever, very large DNA is difficult to isolate intact. During handling, it is randomly sheared to frag-ments in the range of 50,000 to 100,000 base pairs.
A B C
Complete Cleavage
Partial Cleavage
Recognition Sites
A
B
C
A
B
A B C
C
A
B C
A B C
B
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Restriction Enzyme Cleavage of DNA
Plasmids and many viral DNAs are circular molecules. If circular DNA con-tains one recognition site for a restriction enzyme, then it will open up to form a linear molecule when cleaved. By contrast, if a linear DNA molecule contains a single recognition site, when cleaved once it will generate two fragments. The size of the fragments produced depends on how far the sites are from each other. If a DNA molecule contains several recognition sites for a restriction enzyme, then under certain experimental conditions, it is possi-ble that certain sites are cleaved and not others. These incompletely cleaved fragments of DNA are called partials. Partials can arise if low amounts of enzyme are used or the reaction is stopped after a short time. In reality,
reactions containing partials also contain some molecules that have been completely cleaved.
Agarose gel electrophoresis is a powerful sepa-ration method used to analyze DNA fragments generated by restriction enzymes. The agarose gel consists of microscopic pores that act as a molecu-lar sieve. Samples of DNA are loaded into wells made in the gel during casting. Since DNA has a strong negative charge at neutral pH, it migrates through the gel towards the positive electrode during electrophoresis. DNA molecules are sepa-rated in the gel according to their size and shape. The smaller linear fragments migrate the fastest. If the size of two fragments are similar or identi-cal, they will migrate together in the gel. If DNA is cleaved many times, the wide range of fragments
produced will appear as a smear after electrophoresis.
Circular DNAs such as plasmids are supercoiled. Supercoiled DNA has a more compact and entangled shape (like a twisted rub-ber band) than its corresponding non-supercoiled forms (linear, nicked and relaxed circles).
When supercoiled DNA is cleaved once by a restriction enzyme, it unravels to its linear form. If supercoiled DNA is nicked (a phosphate bond is cleaved anywhere in the molecule, in either strand) it completely unravels to a circular form. Under the electrophoresis conditions being used in this experiment, super-coiled DNA migrates faster than its linear form and linear DNA migrates faster than its nicked circular form. During replica-tion, several plasmid molecules can form interlocking structures. These forms are called catenanes. Catenanes can contain two plasmid molecules (dimer), three molecules (trimer), etc. Cat-enanes migrate more slowly than single circles that are nicked during electrophoresis. Dimers migrate faster than trimers, which migrate faster than tetramers, etc.
Dimer
"Nicks" will convert superhelical DNA
Trimer
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Restriction Enzyme Cleavage of DNA
Catenanes give rise to the same final restriction enzyme cleavage patterns as their uncatenated single forms.
In this experiment, restriction enzyme cleavage products will be analyzed by agarose gel electrophoresis. The supercoiled plasmid DNA contains approxi-mately 4,500 base pairs and has one recognition site for Bgl I and two for Eco RI. The second DNA is isolated from the E. coli bacteriophage lambda, which is a linear molecule containing 49,000 base pairs. Lambda DNA con-tains 5 recognition sites for Eco RI and 29 for Bgl I. The restriction enzyme digestions demonstrate that a specific restriction enzyme will yield distinctly different patterns when digesting different DNAs. For example, Bgl I will give one fragment when digesting the plasmid DNA, compared to a signifi-cantly different pattern when digesting lambda DNA.
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Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/labora-tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1989,1992,1994,1997,1998, 2000, 2004, 2007, EDVOTEK, Inc., all rights reserved. EVT 001197AM
The Exp
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ExpERImENT OBjECTIVE:
The objective of this experiment module is to develop an understanding of the use of restriction endonucleases as tools to cut DNA at specific sequences.
LABORATORy sAFETy
1. Gloves and goggles should be worn routinely as good laboratory prac-tice.
2. Exercise extreme caution when working with equipment that is used in conjunction with the heating and/or melting of reagents.
3. DO NOT MOUTH PIPET REAGENTS - USE PIPET PUMPS.
4. Exercise caution when using any electrical equipment in the laboratory.
5. Always wash hands thoroughly with soap and water after handling reagents or biologi-cal materials in the laboratory.
LABORATORy NOTEBOOK RECORDINGs:
Address and record the following in your labora-tory notebook or on a separate worksheet.
Before starting the Experiment:
• Write a hypothesis that reflects the experiment. • Predict experimental outcomes.
During the Experiment:
• Record (draw) your observations, or photograph the results.
Following the Experiment:
• Formulate an explanation from the results. • Determine what could be changed in the experiment if the experi-
ment were repeated. • Write a hypothesis that would reflect this change.
Experiment Overview and General Instructions
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/labora-tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1989,1992,1994,1997,1998, 2000, 2004, 2007, EDVOTEK, Inc., all rights reserved. EVT 001197AM
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DNA Samples
After electrophoresis, transfer gel for staining
Analysis on UV Transilluminator
No destaining
Analysis on White light source After destaining
100 Series (Standard)
InstaStain® Methylene Blue
100-Q SeriesInstaStain®
Ethidium Bromide
© 2
006
EDV
OTE
K, I
nc.
Attach safety cover, connect leads to power source and
conduct electrophoresis
Load each samplein consecutive wells.
Remove end blocks, comb and submerge gel under buffer
in electrophoresis chamber
Prepare agarosegel in casting tray
Gel pattern will vary depending on experiment
Experiment Overview and General Instructions
ExpERImENT OVERVIEw: FLOw ChART
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The Exp
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Individual 1.5 ml or 0.5 ml microtest tubes
• Your instructor may have aliquoted samples into a set of tubes for each lab group. Alternatively, you may be required to withdraw the appropri-ate amount of sample from the experiment stock tubes.
• Check the sample volume. Sometimes a small amount of sample will cling to the walls of the tubes. Make sure the entire volume of sample is at the bottom of the tubes before starting to load the gel.
• Briefly centrifuge the sample tubes, or tap each tube on the table-top to get all the sample to the bottom of the tube.
Experiment Overview and General Instructions
ABOuT ThE ELECTROphOREsIs sAmpLEs
Samples in EDVOTEK Series 100, 100-Q and Sci-On® Series electrophoresis experiments are packaged in individual 1.5 ml or 0.5 ml microtest tubes.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/labora-tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1989,1992,1994,1997,1998, 2000, 2004, 2007, EDVOTEK, Inc., all rights reserved. EVT 001197AM
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AGAROsE GEL REQuIREmENTs FOR ThIs ExpERImENT
• Recommended gel size: 7 x 7 cm or 7 x 14 cm
• Number of sample wells required: 6
• Placement of well-former template: first set of notches
• Agarose gel concentration: 0.8%
pREpARING ThE GEL BED
1. Close off the open ends of a clean and dry gel bed (casting tray) by us-ing rubber dams or tape.
A. Using Rubber dams:
• Place a rubber dam on each end of the bed. Make sure the dam fits firmly in contact with the sides and bottom of the bed.
B. Taping with labeling or masking tape:
• With 3/4 inch wide tape, extend the tape over the sides and bottom edge of the bed.
• Fold the extended edges of the tape back onto the sides and bottom. Press contact points firmly to form a good seal.
2. Place a well-former template (comb) in the first set of notches at the end of the bed. Make sure the comb sits firmly and evenly across the bed.
Agarose Gel Electrophoresis
• If you will be staining the gel after electro-phoresis with InstaStain® Methylene Blue, you should be using Table A.1 or A.2 found in Appendix A.
• If you will be staining the gel after electro-phoresis with InstaStain® Ethidium Bromide, you should be using Table A.3 or A.4 found in Appendix B.
CAsTING AGAROsE GELs
3. Use a 250 ml flask or beaker to prepare the gel solution.
4. Use the appropriate Reference Table for aga-rose gel preparation provided by your instruc-tor. Add the specified amount of agarose powder and buffer and swirl the mixture to disperse clumps of agarose powder.
5. With a marking pen, indicate the level of the solution volume on the outside of the flask.
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The Exp
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6. Heat the mixture to dissolve the agarose powder.
A. Microwave method: • Cover the flask with plastic wrap to minimize evaporation. • Heat the mixture on High for 1 minute. • Swirl the mixture and heat on High in bursts of 25 seconds until
all the agarose is completely dissolved.
B. Hot plate method: • Cover the flask with aluminum foil to minimize evaporation. • Heat the mixture to boiling over a burner with occasional swirl-
ing. Boil until all the agarose is completely dissolved.
Check the solution carefully and continue heating until the final solu-tion appears clear (like water). If "crystal" particles are visible, the agarose is not completely dissolved.
7. Cool the agarose solution to 60°C with careful swirling to promote even dissipation of heat. If detectable evaporation has occurred, add distilled water to bring the solution up to the original volume marked in step 5.
After the gel is cooled to �0°C:
• If you are using rubber dams, go to step 9. • If you are using tape, continue with step 8.
8. Seal the interface of the gel bed and tape to prevent the agarose solution from leaking.
• Use a transfer pipet to deposit a small amount of cooled agarose to both inside ends of the bed.
• Wait approximately 1 minute for the agarose to solidify.
9. Place the bed on a level surface and pour the cooled (60°C) agarose solu-tion into the bed.
10. Allow the gel to completely solidify. It will become firm and cool to the touch after approximately 20 minutes.
pREpARING ThE GEL FOR ELECTROphOREsIs
11. After the gel is completely solidified, carefully and slowly remove the rubber dams or tape from the gel bed.
Be especially careful not to damage or tear the gel wells when removing the rubber dams. A thin plastic knife, spatula or pipet tip can be inserted between the gel and the dams to break the surface tension.
At high altitudes, it is recommended to use a microwave oven to reach boiling temperatures.
Agarose Gel Electrophoresis
DO NOT POUR BOILING HOT AGAROSE INTO THE GEL BED.
Hot agarose solution may irreversibly warp the bed.
60˚C
Important Note
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12. Remove the comb by slowly pulling straight up. Do this carefully and evenly to prevent tearing the sample wells.
13. Place the gel (on its bed) into the electrophoresis chamber, properly oriented, centered and level on the platform.
14. Fill the electrophoresis apparatus chamber with the appropriate amount of diluted (1x) electrophoresis buffer (refer to Table B (found in Appendix A or B) on the instruction sheet provided by your instructor).
15. Make sure that the gel is completely submerged under buffer before proceeding to loading the samples and conducting electrophoresis.
Agarose Gel Electrophoresis
LOADING ThE sAmpLEs
Samples should be loaded into the wells of the gel in consecutive order.
• For gels to be stained with InstaStain® Methylene blue, the amount of sample that should be loaded is 35-38 µl.
• For gels to be stained with InstaStain® Ethidium Bromide, the amount of sample that should be loaded is 18-20 µl.
Reminder:
During electrophoresis, the DNA samples migrate through the agarose gel towards the positive electrode. Before loading the samples, make sure the gel is properly oriented in the apparatus chamber.
+Black Red
Sample wells
–
Lane Tube
1 A Plasmid DNA (uncut) 2 B Plasmid cut with Bgl l 3 C Plasmid cut with Eco Rl 4 D Lambda DNA (uncut) 5 E Lambda DNA cut with Eco Rl 6 F Lambda DNA cut with Bgl l
15Restriction Enzyme Cleavage of DNA
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The Exp
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Agarose Gel Electrophoresis
RuNNING ThE GEL
1. After the DNA samples are loaded, carefully snap the cover down onto the electrode terminals. Make sure that the negative and positive color-coded indicators on the cover and apparatus chamber are properly oriented.
2. Insert the plug of the black wire into the black input of the power source (negative input). Insert the plug of the red wire into the red input of the power source (positive input).
3. Set the power source at the required voltage and conduct electrophore-sis for the length of time determined by your instructor.
4. Check to see that current is flowing properly - you should see bubbles forming on the two platinum electrodes.
5. After the electrophoresis is completed, turn off the power, unplug the power source, disconnect the leads and remove the cover.
6. Remove the gel from the bed for staining.
sTAINING AND VIsuALIzATION OF DNA After electrophoresis, agarose gels require staining to visualize the separated DNA samples. Various options are available for DNA staining. Your instruc-tor will provide instructions for the DNA staining method you will be using.
InstaStain is a registered trademark of EDVOTEK, Inc. Patents Pending.
Electrophoresis can be completed in 15-20 minutes under optimal conditions. For Time and Voltage recommendations, refer to Table C (in Appendix A or B).
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/labora-tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the written consent of EDVOTEK, Inc. Copyright © 1989,1992,1994,1997,1998, 2000, 2004, 2007, EDVOTEK, Inc., all rights reserved. EVT 001197AM
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102Restriction Enzyme Cleavage of DNA
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Answer the following study questions in your laboratory notebook or on a separate worksheet.
1. To which electrode does DNA migrate and why?
2. Why has the discovery of restriction enzymes been so important?
3. How are restriction enzymes named?
4. Briefly explain the function of restriction enzymes.
study Questions
material safety Data sheetsFull size (8.5 x 11”) pdf copy of MSDS available at www.edvotek.com or by request.
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ber
fo
r in
form
atio
n
Dat
e Pr
epar
ed
Sig
nat
ure
of
Prep
arer
(o
pti
on
al)
Ad
dre
ss (
Nu
mb
er, S
tree
t, C
ity,
Sta
te,
Zip
Co
de)
EDV
OTE
K, I
nc.
1467
6 R
oth
geb
Dri
veR
ock
ville
, MD
208
50
Haz
ard
ou
s C
om
po
nen
ts [
Spec
ific
C
hem
ical
Iden
tity
; C
om
mo
n N
ame(
s)]
O
SHA
PEL
AC
GIH
TLV
Oth
er L
imit
s R
eco
mm
end
ed%
(O
pti
on
al)
(301
) 25
1-59
90
(301
) 25
1-59
90
Bo
ilin
g P
oin
t
Sect
ion
III -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
s
Un
usu
al F
ire
and
Exp
losi
on
Haz
ard
s
Spec
ial F
ire
Fig
hti
ng
Pro
ced
ure
s
Vap
or
Pres
sure
(m
m H
g.)
Vap
or
Den
sity
(A
IR =
1)
Solu
bili
ty in
Wat
er
Ap
pea
ran
ce a
nd
Od
or
Sect
ion
IV -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
sFl
ash
Po
int
(Met
ho
d U
sed
)
Exti
ng
uis
hin
g M
edia
Flam
mab
le L
imit
sU
ELLE
L
Mel
tin
g P
oin
t
Evap
ora
tio
n R
ate
(Bu
tyl A
ceta
te =
1)
Spec
ific
Gra
vity
(H
0 =
1)
2
Ag
aro
se
10/0
5/06
This
pro
du
ct c
on
tain
s n
o h
azar
do
us
mat
eria
ls a
s d
efin
ed b
y th
e O
SHA
Haz
ard
Co
mm
un
icat
ion
Stan
dar
d.
CA
S #9
012-
36-6
For
1% s
olu
tio
n 1
94 F
N
o d
ata
N
o d
ata
No
dat
a
No
dat
a
No
dat
a
Inso
lub
le -
co
ld
W
hit
e p
ow
der
, no
od
or
N.D
. = N
o d
ata
No
dat
a
N
.D.
N.D
.
Wat
er s
pra
y, d
ry c
hem
ical
, car
bo
n d
ioxi
de,
hal
on
or
stan
dar
d f
oam
Poss
ible
fir
e h
azar
d w
hen
exp
ose
d t
o h
eat
or
flam
e
No
ne
ED
VO
TE
K®
Stab
ility
Sect
ion
V -
Rea
ctiv
ity
Dat
aU
nst
able
Sect
ion
VI -
Hea
lth
Haz
ard
Dat
a
Inco
mp
atib
ility
Co
nd
itio
ns
to A
void
Ro
ute
(s)
of
Entr
y:In
hal
atio
n?
Ing
esti
on
?Sk
in?
Oth
er
Stab
le
Haz
ard
ou
s Po
lym
eriz
atio
nM
ay O
ccu
rC
on
dit
ion
s to
Avo
id
Will
No
t O
ccu
r
Hea
lth
Haz
ard
s (A
cute
an
d C
hro
nic
)
Car
cin
og
enic
ity:
NTP
?O
SHA
Reg
ula
tio
n?
IAR
C M
on
og
rap
hs?
Sig
ns
and
Sym
pto
ms
of
Exp
osu
re
Med
ical
Co
nd
itio
ns
Gen
eral
ly A
gg
rava
ted
by
Exp
osu
re
Emer
gen
cy F
irst
Aid
Pro
ced
ure
s
Sect
ion
VII
- Pr
ecau
tio
ns
for
Safe
Han
dlin
g a
nd
Use
Step
s to
be
Take
n in
cas
e M
ater
ial i
s R
elea
sed
fo
r Sp
illed
Was
te D
isp
osa
l Met
ho
d
Prec
auti
on
s to
be
Take
n in
Han
dlin
g a
nd
Sto
rin
g
Oth
er P
reca
uti
on
s
Sect
ion
VIII
- C
on
tro
l Mea
sure
s
Ven
tila
tio
nLo
cal E
xhau
stSp
ecia
l
Mec
han
ical
G
en. d
iluti
on
ven
tila
tio
n
Res
pir
ato
ry P
rote
ctio
n (
Spec
ify
Typ
e)
Pro
tect
ive
Glo
ves
Oth
er P
rote
ctiv
e C
loth
ing
or
Equ
ipm
ent
Wo
rk/H
ygie
nic
Pra
ctic
es
Eye
Pro
tect
ion
Haz
ard
ou
s D
eco
mp
osi
tio
n o
r B
ypro
du
cts
Yes
Sp
lash
pro
of
go
gg
les
Imp
ervi
ou
s cl
oth
ing
to
pre
ven
t sk
in c
on
tact
No
neX
N
on
e
No
dat
a av
aila
ble
X
No
ne
Yes
Y
es
Yes
Inh
alat
ion
: N
o d
ata
avai
lab
le
In
ges
tio
n:
Larg
e am
ou
nts
may
cau
se d
iarr
hea
No
dat
a av
aila
ble
No
dat
a av
aila
ble
Trea
t sy
mp
tom
atic
ally
an
d s
up
po
rtiv
ely
Swee
p u
p a
nd
pla
ce in
su
itab
le c
on
tain
er f
or
dis
po
sal
No
rmal
so
lid w
aste
dis
po
sal
No
ne
No
ne
Ch
emic
al c
artr
idg
e re
spir
ato
r w
ith
fu
ll fa
cep
iece
.
ED
VO
TE
K®
Mat
eria
l Saf
ety
Dat
a Sh
eet
May
be
use
d t
o c
om
ply
wit
h O
SHA
's H
azar
d C
om
mu
nic
atio
nSt
and
ard
. 29
CFR
191
0.12
00 S
tan
dar
d m
ust
be
con
sult
ed f
or
spec
ific
req
uir
emen
ts.
IDEN
TITY
(A
s U
sed
on
Lab
el a
nd
Lis
t)N
ote
: B
lan
k sp
aces
are
no
t p
erm
itte
d.
If a
ny
item
is n
ot
app
licab
le, o
r n
o in
form
atio
n is
ava
ilab
le, t
he
spac
e m
ust
b
e m
arke
d t
o in
dic
ate
that
.
Sect
ion
IM
anu
fact
ure
r's
Nam
e
Sect
ion
II -
Haz
ard
ou
s In
gre
die
nts
/Id
enti
fy In
form
atio
n
Emer
gen
cy T
elep
ho
ne
Nu
mb
er
Tele
ph
on
e N
um
ber
fo
r in
form
atio
n
Dat
e Pr
epar
ed
Sig
nat
ure
of
Prep
arer
(o
pti
on
al)
Ad
dre
ss (
Nu
mb
er, S
tree
t, C
ity,
Sta
te,
Zip
Co
de)
EDV
OTE
K, I
nc.
1467
6 R
oth
geb
Dri
veR
ock
ville
, MD
208
50
Haz
ard
ou
s C
om
po
nen
ts [
Spec
ific
C
hem
ical
Iden
tity
; C
om
mo
n N
ame(
s)]
O
SHA
PEL
AC
GIH
TLV
Oth
er L
imit
s R
eco
mm
end
ed%
(O
pti
on
al)
(301
) 25
1-59
90
(301
) 25
1-59
90
Bo
ilin
g P
oin
t
Sect
ion
III -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
s
Un
usu
al F
ire
and
Exp
losi
on
Haz
ard
s
Spec
ial F
ire
Fig
hti
ng
Pro
ced
ure
s
Vap
or
Pres
sure
(m
m H
g.)
Vap
or
Den
sity
(A
IR =
1)
Solu
bili
ty in
Wat
er
Ap
pea
ran
ce a
nd
Od
or
Sect
ion
IV -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
sFl
ash
Po
int
(Met
ho
d U
sed
)
Exti
ng
uis
hin
g M
edia
Flam
mab
le L
imit
sU
ELLE
L
Mel
tin
g P
oin
t
Evap
ora
tio
n R
ate
(Bu
tyl A
ceta
te =
1)
Spec
ific
Gra
vity
(H
0 =
1)
2
50x
Elec
tro
ph
ore
sis
Bu
ffer
This
pro
du
ct c
on
tain
s n
o h
azar
do
us
mat
eria
ls a
s d
efin
ed b
y th
e O
SHA
Haz
ard
Co
mm
un
icat
ion
Sta
nd
ard
.
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
Ap
pre
ciab
le, (
gre
ater
th
an 1
0%)
Cle
ar, l
iqu
id, s
ligh
t vi
neg
ar o
do
r
No
dat
a
N.D
. = N
o d
ata N.D
.
N.D
.
Use
ext
ing
uis
hin
g m
edia
ap
pro
pri
ate
for
surr
ou
nd
ing
fir
e.
Wea
r p
rote
ctiv
e eq
uip
men
t an
d S
CB
A w
ith
fu
ll fa
cep
iece
op
erat
ed in
po
siti
ve p
ress
ure
mo
de.
No
ne
iden
tifi
ed
10/0
5/06
Stab
ility
Sect
ion
V -
Rea
ctiv
ity
Dat
aU
nst
able
Sect
ion
VI -
Hea
lth
Haz
ard
Dat
a
Inco
mp
atib
ility
Co
nd
itio
ns
to A
void
Ro
ute
(s)
of
Entr
y:In
hal
atio
n?
Ing
esti
on
?Sk
in?
Oth
er
Stab
le
Haz
ard
ou
s Po
lym
eriz
atio
nM
ay O
ccu
rC
on
dit
ion
s to
Avo
id
Will
No
t O
ccu
r
Hea
lth
Haz
ard
s (A
cute
an
d C
hro
nic
)
Car
cin
og
enic
ity:
NTP
?O
SHA
Reg
ula
tio
n?
IAR
C M
on
og
rap
hs?
Sig
ns
and
Sym
pto
ms
of
Exp
osu
re
Med
ical
Co
nd
itio
ns
Gen
eral
ly A
gg
rava
ted
by
Exp
osu
re
Emer
gen
cy F
irst
Aid
Pro
ced
ure
s
Sect
ion
VII
- Pr
ecau
tio
ns
for
Safe
Han
dlin
g a
nd
Use
Step
s to
be
Take
n in
cas
e M
ater
ial i
s R
elea
sed
fo
r Sp
illed
Was
te D
isp
osa
l Met
ho
d
Prec
auti
on
s to
be
Take
n in
Han
dlin
g a
nd
Sto
rin
g
Oth
er P
reca
uti
on
s
Sect
ion
VIII
- C
on
tro
l Mea
sure
s
Ven
tila
tio
nLo
cal E
xhau
stSp
ecia
l
Mec
han
ical
(G
ener
al)
Res
pir
ato
ry P
rote
ctio
n (
Spec
ify
Typ
e)
Pro
tect
ive
Glo
ves
Oth
er P
rote
ctiv
e C
loth
ing
or
Equ
ipm
ent
Wo
rk/H
ygie
nic
Pra
ctic
es
Eye
Pro
tect
ion
Haz
ard
ou
s D
eco
mp
osi
tio
n o
r B
ypro
du
cts
X
N
on
e
Stro
ng
oxi
diz
ing
ag
ents
Car
bo
n m
on
oxi
de,
Car
bo
n d
ioxi
de
X
N
on
e
Yes
Y
es
Y
es
No
ne
No
ne
iden
tifi
ed
Irri
tati
on
to
up
per
res
pir
ato
ry t
ract
, ski
n, e
yes
No
ne
Ing
esti
on
: If
co
nsc
iou
s, g
ive
larg
e am
ou
nts
of
wat
er
Eyes
: Fl
ush
wit
h w
ater
In
hal
atio
n:
Mo
ve t
o f
resh
air
Sk
in:
Was
h w
ith
so
ap a
nd
wat
er
Wea
r su
itab
le p
rote
ctiv
e cl
oth
ing
. M
op
up
sp
ill
and
rin
se w
ith
wat
er, o
r co
llect
in a
bso
rpti
ve m
ater
ial a
nd
dis
po
se o
f th
e ab
sorp
tive
mat
eria
l.
Dis
po
se in
acc
ord
ance
wit
h a
ll ap
plic
able
fed
eral
, sta
te, a
nd
loca
l en
viro
men
tal r
egu
lati
on
s.
Avo
id e
ye a
nd
ski
n c
on
tact
.
No
ne
Yes
N
on
e
Yes
N
on
e
Yes
_Saf
ety
go
gg
les
No
ne
No
ne
Stab
ility
Sect
ion
V -
Rea
ctiv
ity
Dat
aU
nst
able
Sect
ion
VI -
Hea
lth
Haz
ard
Dat
a
Inco
mp
atib
ility
Co
nd
itio
ns
to A
void
Ro
ute
(s)
of
Entr
y:In
hal
atio
n?
Ing
esti
on
?Sk
in?
Oth
er
Stab
le
Haz
ard
ou
s Po
lym
eriz
atio
nM
ay O
ccu
rC
on
dit
ion
s to
Avo
id
Will
No
t O
ccu
r
Hea
lth
Haz
ard
s (A
cute
an
d C
hro
nic
)
Car
cin
og
enic
ity:
NTP
?O
SHA
Reg
ula
tio
n?
IAR
C M
on
og
rap
hs?
Sig
ns
and
Sym
pto
ms
of
Exp
osu
re
Med
ical
Co
nd
itio
ns
Gen
eral
ly A
gg
rava
ted
by
Exp
osu
re
Emer
gen
cy F
irst
Aid
Pro
ced
ure
s
Sect
ion
VII
- Pr
ecau
tio
ns
for
Safe
Han
dlin
g a
nd
Use
Step
s to
be
Take
n in
cas
e M
ater
ial i
s R
elea
sed
fo
r Sp
illed
Was
te D
isp
osa
l Met
ho
d
Prec
auti
on
s to
be
Take
n in
Han
dlin
g a
nd
Sto
rin
g
Oth
er P
reca
uti
on
s
Sect
ion
VIII
- C
on
tro
l Mea
sure
s
Ven
tila
tio
nLo
cal E
xhau
stSp
ecia
l
Mec
han
ical
(G
ener
al)
Res
pir
ato
ry P
rote
ctio
n (
Spec
ify
Typ
e)
Pro
tect
ive
Glo
ves
Oth
er P
rote
ctiv
e C
loth
ing
or
Equ
ipm
ent
Wo
rk/H
ygie
nic
Pra
ctic
es
Eye
Pro
tect
ion
Haz
ard
ou
s D
eco
mp
osi
tio
n o
r B
ypro
du
cts
X
N
on
e
Stro
ng
oxi
diz
ing
ag
ents
Car
bo
n m
on
oxi
de,
Car
bo
n d
ioxi
de,
nit
rog
en o
xid
es, h
ydro
gen
bro
mid
e g
as
X
N
on
e
Yes
Y
es
Yes
No
dat
a av
aila
ble
Irri
tati
on
to
mu
cou
s m
emb
ran
es a
nd
up
per
res
pir
ato
ry t
ract
No
dat
a
Trea
t sy
mp
tom
atic
ally
an
d s
up
po
rtiv
ely
Wea
r SC
BA
, ru
bb
er b
oo
ts, r
ub
ber
glo
ves
Mix
mat
eria
l wit
h c
om
bu
stib
le s
olv
ent
and
bu
rn in
a c
hem
ical
inci
ner
ato
r eq
uip
ped
aft
erb
urn
er a
nd
scr
ub
ber
Use
in c
hem
ical
fu
me
ho
od
wit
h p
rop
er p
rote
ctiv
e la
b g
ear.
Mu
tag
en
Yes
Ch
em. f
um
e h
oo
d
No
N
on
e
Ru
bb
er
C
hem
. saf
ety
go
gg
les
R
ub
ber
bo
ots
Use
in c
hem
ical
fu
me
ho
od
wit
h p
rop
er p
rote
ctiv
e la
b g
ear.
Acu
te: M
ater
ial i
rrit
atin
g t
o m
uco
us
mem
bra
nes
, up
per
res
pir
ato
ry t
ract
, eye
s, s
kin
Ch
ron
ic:
May
alt
er g
enet
ic m
ater
ial
SCB
A
Mat
eria
l Saf
ety
Dat
a Sh
eet
May
be
use
d t
o c
om
ply
wit
h O
SHA
's H
azar
d C
om
mu
nic
atio
nSt
and
ard
. 29
CFR
191
0.12
00 S
tan
dar
d m
ust
be
con
sult
ed f
or
spec
ific
req
uir
emen
ts.
IDEN
TITY
(A
s U
sed
on
Lab
el a
nd
Lis
t)N
ote
: B
lan
k sp
aces
are
no
t p
erm
itte
d.
If a
ny
item
is n
ot
app
licab
le, o
r n
o in
form
atio
n is
ava
ilab
le, t
he
spac
e m
ust
b
e m
arke
d t
o in
dic
ate
that
.
Sect
ion
IM
anu
fact
ure
r's
Nam
e
Sect
ion
II -
Haz
ard
ou
s In
gre
die
nts
/Id
enti
fy In
form
atio
n
Emer
gen
cy T
elep
ho
ne
Nu
mb
er
Tele
ph
on
e N
um
ber
fo
r in
form
atio
n
Dat
e Pr
epar
ed
Sig
nat
ure
of
Prep
arer
(o
pti
on
al)
Inst
aSta
in, I
nc.
P.O
. Bo
x 12
32W
est
Bet
hes
da,
MD
208
27
Haz
ard
ou
s C
om
po
nen
ts [
Spec
ific
C
hem
ical
Iden
tity
; C
om
mo
n N
ame(
s)]
O
SHA
PEL
AC
GIH
TLV
Oth
er L
imit
s R
eco
mm
end
ed%
(O
pti
on
al)
(301
) 25
1-59
90
(301
) 25
1-59
90
Bo
ilin
g P
oin
t
Sect
ion
III -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
s
Un
usu
al F
ire
and
Exp
losi
on
Haz
ard
s
Spec
ial F
ire
Fig
hti
ng
Pro
ced
ure
s
Vap
or
Pres
sure
(m
m H
g.)
Vap
or
Den
sity
(A
IR =
1)
Solu
bili
ty in
Wat
er
Ap
pea
ran
ce a
nd
Od
or
Sect
ion
IV -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
sFl
ash
Po
int
(Met
ho
d U
sed
)
Exti
ng
uis
hin
g M
edia
Flam
mab
le L
imit
sU
ELLE
L
Mel
tin
g P
oin
t
Evap
ora
tio
n R
ate
(Bu
tyl A
ceta
te =
1)
Spec
ific
Gra
vity
(H
0 =
1)
2
Inst
aSta
in®
Eth
idiu
m B
rom
ide
Eth
idiu
m B
rom
ide
D
ata
no
t av
aila
ble
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
Solu
ble
Ch
emic
al b
ou
nd
to
pap
er, n
o o
do
r
No
dat
a
N.D
. = N
o d
ata N.D
. N
.D.
Wat
er s
pra
y, c
arb
on
dio
xid
e, d
ry c
hem
ical
po
wd
er, a
lco
ho
l or
po
lym
er f
oam
Wea
r p
rote
ctiv
e cl
oth
ing
an
d S
CB
A t
o p
reve
nt
con
tact
wit
h s
kin
& e
yes
Emit
s to
xic
fum
es
10/0
5/06
CA
S# 1
39-3
3-3
(2,7
-Dia
min
o-1
0-Et
hyl
-9-P
hen
ylp
hen
anth
rid
iniu
m B
rom
ide)
ED
VO
TE
K®
material safety Data sheetsFull size (8.5 x 11”) pdf copy of MSDS available at www.edvotek.com or by request.
Mat
eria
l Saf
ety
Dat
a Sh
eet
May
be
use
d t
o c
om
ply
wit
h O
SHA
's H
azar
d C
om
mu
nic
atio
nSt
and
ard
. 29
CFR
191
0.12
00 S
tan
dar
d m
ust
be
con
sult
ed f
or
spec
ific
req
uir
emen
ts.
IDEN
TITY
(A
s U
sed
on
Lab
el a
nd
Lis
t)N
ote
: B
lan
k sp
aces
are
no
t p
erm
itte
d.
If a
ny
item
is n
ot
app
licab
le, o
r n
o in
form
atio
n is
ava
ilab
le, t
he
spac
e m
ust
b
e m
arke
d t
o in
dic
ate
that
.
Sect
ion
IM
anu
fact
ure
r's
Nam
e
Sect
ion
II -
Haz
ard
ou
s In
gre
die
nts
/Iden
tify
Info
rmat
ion
Emer
gen
cy T
elep
ho
ne
Nu
mb
er
Tele
ph
on
e N
um
ber
fo
r in
form
atio
n
Dat
e Pr
epar
ed
Sig
nat
ure
of
Prep
arer
(o
pti
on
al)
Ad
dre
ss (
Nu
mb
er, S
tree
t, C
ity,
Sta
te,
Zip
Co
de)
EDV
OTE
K, I
nc.
1467
6 R
oth
geb
Dri
veR
ock
ville
, MD
208
50
Haz
ard
ou
s C
om
po
nen
ts [
Spec
ific
C
hem
ical
Iden
tity
; C
om
mo
n N
ame(
s)]
O
SHA
PEL
AC
GIH
TLV
Oth
er L
imit
s R
eco
mm
end
ed%
(O
pti
on
al)
(301
) 25
1-59
90
(301
) 25
1-59
90
Bo
ilin
g P
oin
t
Sect
ion
III -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
s
Un
usu
al F
ire
and
Exp
losi
on
Haz
ard
s
Spec
ial F
ire
Fig
hti
ng
Pro
ced
ure
s
Vap
or
Pres
sure
(m
m H
g.)
Vap
or
Den
sity
(A
IR =
1)
Solu
bili
ty in
Wat
er
Ap
pea
ran
ce a
nd
Od
or
Sect
ion
IV -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
sFl
ash
Po
int
(Met
ho
d U
sed
)
Exti
ng
uis
hin
g M
edia
Flam
mab
le L
imit
sU
ELLE
L
Mel
tin
g P
oin
t
Evap
ora
tio
n R
ate
(Bu
tyl A
ceta
te =
1)
Spec
ific
Gra
vity
(H
0 =
1)
2
ED
VO
TE
K®
Inst
aSta
in®
Met
hyl
ene
Blu
e, M
eth
ylen
e B
lue
Plu
s™
10/0
5/06
Met
hyl
ene
Blu
e
3.7
Bis
(D
imet
hyl
amin
o)
Phen
oth
iazi
n 5
IUM
C
hlo
rid
e
No
dat
a av
aila
ble
CA
S #
61-7
3-4
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
Solu
ble
- c
old
Ch
emic
al b
ou
nd
to
pap
er, n
o o
do
r
No
dat
a av
aila
ble
No
dat
a
N
o d
ata
Wat
er s
pra
y, c
arb
on
dio
xid
e, d
ry c
hem
ical
po
wd
er, a
lco
ho
l or
po
lym
er f
oam
Self
co
nta
ined
bre
ath
ing
ap
par
atu
s an
d p
rote
ctiv
e cl
oth
ing
to
pre
ven
t co
nta
ct
wit
h s
kin
an
d e
yes
Emit
s to
xid
fu
mes
un
der
fir
e co
nd
itio
ns
Stab
ility
Sect
ion
V -
Rea
ctiv
ity
Dat
aU
nst
able
Sect
ion
VI -
Hea
lth
Haz
ard
Dat
a
Inco
mp
atib
ility
Co
nd
itio
ns
to A
void
Ro
ute
(s)
of
Entr
y:In
hal
atio
n?
Ing
esti
on
?Sk
in?
Oth
er
Stab
le
Haz
ard
ou
s Po
lym
eriz
atio
nM
ay O
ccu
rC
on
dit
ion
s to
Avo
id
Will
No
t O
ccu
r
Hea
lth
Haz
ard
s (A
cute
an
d C
hro
nic
)
Car
cin
og
enic
ity:
NTP
?O
SHA
Reg
ula
tio
n?
IAR
C M
on
og
rap
hs?
Sig
ns
and
Sym
pto
ms
of
Exp
osu
re
Med
ical
Co
nd
itio
ns
Gen
eral
ly A
gg
rava
ted
by
Exp
osu
re
Emer
gen
cy F
irst
Aid
Pro
ced
ure
s
Sect
ion
VII
- Pr
ecau
tio
ns
for
Safe
Han
dlin
g a
nd
Use
Step
s to
be
Take
n in
cas
e M
ater
ial i
s R
elea
sed
fo
r Sp
illed
Was
te D
isp
osa
l Met
ho
d
Prec
auti
on
s to
be
Take
n in
Han
dlin
g a
nd
Sto
rin
g
Oth
er P
reca
uti
on
s
Sect
ion
VIII
- C
on
tro
l Mea
sure
s
Ven
tila
tio
nLo
cal E
xhau
stSp
ecia
l
Mec
han
ical
(G
ener
al)
Res
pir
ato
ry P
rote
ctio
n (
Spec
ify
Typ
e)
Pro
tect
ive
Glo
ves
Oth
er P
rote
ctiv
e C
loth
ing
or
Equ
ipm
ent
Wo
rk/H
ygie
nic
Pra
ctic
es
Eye
Pro
tect
ion
Haz
ard
ou
s D
eco
mp
osi
tio
n o
r B
ypro
du
cts
X
No
ne
Stro
ng
oxi
diz
ing
ag
ents
Toxi
c fu
mes
of
Car
bo
n m
on
oxi
de,
Car
bo
n d
ioxi
de,
n
itro
gen
oxi
des
, su
lfu
r o
xid
es, h
ydro
gen
, ch
lori
de
gas
X
N
on
e
Yes
Y
es
Yes
Skin
: M
ay c
ause
ski
n ir
rita
tio
n
Eyes
: M
ay c
ause
eye
irri
tati
on
In
hal
atio
n:
Cya
no
sis
Mee
ts c
rite
ria
for
pro
po
sed
OSH
A m
edic
al r
eco
rds
rule
PER
EAC
47.
3042
0.82
No
dat
a av
aila
ble
No
dat
a av
aila
ble
Trea
t sy
mp
tom
atic
ally
Ven
tila
te a
rea
and
was
h s
pill
sit
e
Mix
mat
eria
l wit
h a
co
mb
ust
ible
so
lven
t an
d b
urn
in c
hem
ical
inci
ner
ato
r eq
uip
ped
wit
h a
fter
bu
rner
an
d s
cru
bb
er.
Ch
eck
loca
l an
d s
tate
reg
ula
tio
ns.
Kee
p t
igh
tly
clo
sed
. St
ore
in c
oo
l, d
ry p
lace
No
ne
MIO
SH/O
SHA
ap
pro
ved
, SC
BA
Req
uir
ed
Ru
bb
erC
hem
. saf
ety
go
gg
les
Ru
bb
er b
oo
ts
Stab
ility
Sect
ion
V -
Rea
ctiv
ity
Dat
aU
nst
able
Sect
ion
VI -
Hea
lth
Haz
ard
Dat
a
Inco
mp
atib
ility
Co
nd
itio
ns
to A
void
Ro
ute
(s)
of
Entr
y:In
hal
atio
n?
Ing
esti
on
?Sk
in?
Oth
er
Stab
le
Haz
ard
ou
s Po
lym
eriz
atio
nM
ay O
ccu
rC
on
dit
ion
s to
Avo
id
Will
No
t O
ccu
r
Hea
lth
Haz
ard
s (A
cute
an
d C
hro
nic
)
Car
cin
og
enic
ity:
NTP
?O
SHA
Reg
ula
tio
n?
IAR
C M
on
og
rap
hs?
Sig
ns
and
Sym
pto
ms
of
Exp
osu
re
Med
ical
Co
nd
itio
ns
Gen
eral
ly A
gg
rava
ted
by
Exp
osu
re
Emer
gen
cy F
irst
Aid
Pro
ced
ure
s
Sect
ion
VII
- Pr
ecau
tio
ns
for
Safe
Han
dlin
g a
nd
Use
Step
s to
be
Take
n in
cas
e M
ater
ial i
s R
elea
sed
fo
r Sp
illed
Was
te D
isp
osa
l Met
ho
d
Prec
auti
on
s to
be
Take
n in
Han
dlin
g a
nd
Sto
rin
g
Oth
er P
reca
uti
on
s
Sect
ion
VIII
- C
on
tro
l Mea
sure
s
Ven
tila
tio
nLo
cal E
xhau
stSp
ecia
l
Mec
han
ical
(G
ener
al)
Res
pir
ato
ry P
rote
ctio
n (
Spec
ify
Typ
e)
Pro
tect
ive
Glo
ves
Oth
er P
rote
ctiv
e C
loth
ing
or
Equ
ipm
ent
Wo
rk/H
ygie
nic
Pra
ctic
es
Eye
Pro
tect
ion
Haz
ard
ou
s D
eco
mp
osi
tio
n o
r B
ypro
du
cts
X
N
on
e
No
ne
Sulf
ur
oxi
des
, an
d b
rom
ides
X
N
on
e
Yes
Y
es
Yes
Acu
te e
ye c
on
tact
: M
ay c
ause
irri
tati
on
. N
o d
ata
avai
lab
le f
or
oth
er r
ou
tes.
No
dat
a av
aila
ble
May
cau
se s
kin
or
eye
irri
tati
on
No
ne
rep
ort
ed
Trea
t sy
mp
tom
atic
ally
an
d s
up
po
rtiv
ely.
Rin
se c
on
tact
ed a
rea
wit
h c
op
iou
s am
ou
nts
of
wat
er.
Wea
r ey
e an
d s
kin
pro
tect
ion
an
d m
op
sp
ill a
rea.
Rin
se w
ith
wat
er.
Ob
serv
e al
l fed
eral
, sta
te, a
nd
loca
l reg
ula
tio
ns.
Avo
id e
ye a
nd
ski
n c
on
tact
.
No
ne
Yes
No
ne
Yes
No
ne
Yes
Spla
sh p
roo
f g
og
gle
s
No
ne
req
uir
ed
Avo
id e
ye a
nd
ski
n c
on
tact
Mat
eria
l Saf
ety
Dat
a Sh
eet
May
be
use
d t
o c
om
ply
wit
h O
SHA
's H
azar
d C
om
mu
nic
atio
nSt
and
ard
. 29
CFR
191
0.12
00 S
tan
dar
d m
ust
be
con
sult
ed f
or
spec
ific
req
uir
emen
ts.
IDEN
TITY
(A
s U
sed
on
Lab
el a
nd
Lis
t)N
ote
: B
lan
k sp
aces
are
no
t p
erm
itte
d.
If a
ny
item
is n
ot
app
licab
le, o
r n
o in
form
atio
n is
ava
ilab
le, t
he
spac
e m
ust
b
e m
arke
d t
o in
dic
ate
that
.
Sect
ion
IM
anu
fact
ure
r's
Nam
e
Sect
ion
II -
Haz
ard
ou
s In
gre
die
nts
/Id
enti
fy In
form
atio
n
Emer
gen
cy T
elep
ho
ne
Nu
mb
er
Tele
ph
on
e N
um
ber
fo
r in
form
atio
n
Dat
e Pr
epar
ed
Sig
nat
ure
of
Prep
arer
(o
pti
on
al)
Ad
dre
ss (
Nu
mb
er, S
tree
t, C
ity,
Sta
te,
Zip
Co
de)
EDV
OTE
K, I
nc.
1467
6 R
oth
geb
Dri
veR
ock
ville
, MD
208
50
Haz
ard
ou
s C
om
po
nen
ts [
Spec
ific
C
hem
ical
Iden
tity
; C
om
mo
n N
ame(
s)]
O
SHA
PEL
AC
GIH
TLV
Oth
er L
imit
s R
eco
mm
end
ed%
(O
pti
on
al)
(301
) 25
1-59
90
(301
) 25
1-59
90
Bo
ilin
g P
oin
t
Sect
ion
III -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
s
Un
usu
al F
ire
and
Exp
losi
on
Haz
ard
s
Spec
ial F
ire
Fig
hti
ng
Pro
ced
ure
s
Vap
or
Pres
sure
(m
m H
g.)
Vap
or
Den
sity
(A
IR =
1)
Solu
bili
ty in
Wat
er
Ap
pea
ran
ce a
nd
Od
or
Sect
ion
IV -
Ph
ysic
al/C
hem
ical
Ch
arac
teri
stic
sFl
ash
Po
int
(Met
ho
d U
sed
)
Exti
ng
uis
hin
g M
edia
Flam
mab
le L
imit
sU
ELLE
L
Mel
tin
g P
oin
t
Evap
ora
tio
n R
ate
(Bu
tyl A
ceta
te =
1)
Spec
ific
Gra
vity
(H
0 =
1)
2
Prac
tice
Gel
Lo
adin
g S
olu
tio
n
10/0
5/06
This
pro
du
ct c
on
tain
s n
o h
azar
do
us
mat
eria
ls a
s d
efin
ed b
y th
e O
SHA
Haz
ard
Co
mm
un
icat
ion
Stan
dar
d.
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
No
dat
a
Solu
ble
B
lue
liqu
id, n
o o
do
r
No
dat
aN
o d
ata
No
dat
a
Dry
ch
emic
al, c
arb
on
dio
xid
e, w
ater
sp
ray
or
foam
Use
age
nts s
uita
ble
for t
ype
of su
rrou
ndin
g fir
e. K
eep
upw
ind,
avo
idb
reat
hin
g h
azar
do
us
sulf
ur
oxi
des
an
d b
rom
ides
. W
ear
SCB
A.
Un
kno
wn
ED
VO
TE
K®