gel documentation systemsuv-light for up to 60 seconds. the pcr product was subsequently isolated...

NIPPON Genetics EUROPE

Gel Documentation Systems

Technology

Why is UV-light so dangerous?

IntroductionDetection of nucleic acid is still performed using light in the UV-range and ethidium bromide as nucleic acid stains. However, the degradation of nucleic acids by UV-light is well documented and can therefore be considered a nucleic acid destroying agent. Additionally, the UV-light is considered dangerous for the user. Overall, the efficiency of downstream applications, such as sequencing, cloning, etc. can be significantly reduced due to damaged DNA molecules. Harmless visible light has already been used to detect nucleic acids with its major limitation of only exciting green dyes. NIPPON Genetics EUROPE, known for its safe nucleic acid dyes MIDORIGreen, has developed a complete new approach for the detection of nucleic acid dyes able to excite all commercially available nucleic acid dyes.

DNA is able to absorb light in the UV-spectrum. Consequences are the formation of pyrimidine dimers; ssDNA and dsDNA breaks, DNA-DNA interstrand crosslinks and DNA-to-protein crosslinks.

Organisms are able to repair these DNA damages or suffer the consequences discussed above. DNA in agarose gels lack these repair mechanisms.

Additionally, the amount of DNA in a gel is much less when compared to a whole organism. Therefore DNA degradation caused by short-term UV light exposure was investigated:

After an electrophoretic run, the gel was exposed to UV-light for up to 60 seconds. The PCR product was subsequently isolated and cloned into a control vector, previously double digested with NdeI and HindIII, before being cloned and transformed into Escherichia coli DH5α (BL21) cells. The results are shown below:

The DNA exposed to UV-light was highly damaged, significantly reducing the cloning efficiency. After 60 seconds exposure, no colony was detected, indicating complete damage of DNA.

In 2009, blue light emitting diodes (LED) transilluminators were introduced. LED are semi-conductors able to transform electricity into light. The advantages are defined wavelengths, superb efficiency and a huge average life expectancy of 50,000 hours. They emit light with a wavelength of 470 nm, which is after the UV-wavelength range.

The experiment described previously was repeated and the DNA was exposed to the blue LED light for the same amount of time:

As expected from the physical theory, the amount of CFU per plate did not have any significant change after the exposure. The prevention of the DNA damage therefore vastly increased the cloning efficiency.

Blue LED - First Solution

Problems with Blue LED

Although having the assets of safe light source, using blue LEDs also had a central drawback. Ethidium bromide, the most common DNA stain did only deliver poor results. The

excitation peak of ethidium bromide bound to DNA is between 480 and 525 nm.

As can be seen, the signal of ethidium bromide bound to DNA is very week and cannot be used. Green DNA dyes, such as MIDORIGreen dyes, deliver a much better signal with blue LEDs.

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NIPPON Genetics EUROPE presents a new strategy for the detection of nucleic acid dyes:

Instead of using a single wavelength, a combination of wavelengths was created resulting in a spectrum of light with blue/green light of 470 nm to 520 nm, able to excite dyes which were not excitable with a single wavelength of 470 nm, such as ethidium bromide. This can be achieved by the accumulated energy absoprtion of the dyes.

This idea led to the development of the FastGene® Blue/Green LED Transilluminator (FG-08): Signal intensities as high as from strong UV-transilluminators using safe wavelength were finally possible for all DNA dyes. Ethidium bromide and the safe alternatives deliver a much better signal with this unique technology, with much better results in downstream applications as well as creating a safer laboratory environment.

NIPPON Genetics EUROPE presents here its stand alone imaging systems and illuminators using this revolutionary technology.

-Technology

For more information or for a free demonstration contact us at [email protected]

Absortion curve of fluorescent dyes attached to DNA

„I have just used a Blue/Green LED illuminator with MIDORIGreen Direct and it never fails to impress!“

Dr. Jean Emly, Founder of Geneflow Ltd. , England

• Mixing the LED wavelengths creates an optimal spectrum of light to excite a great variety of dyes.

• No degradation of DNA/RNA

• Ethidium bromide produces great signal

• Also able to excite fluorescent proteins, such as GFP, RFP, etc.

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- The Revolution

MIDORIGreen Direct

MIDORIGreen Advance MIDORIGreen Advance

Ethidium bromideEthidium bromide

MIDORIGreen DirectUV-LightB/G LED-Light

Imaging Systems

GelPic LED Box GelPicBox

Cat. No. GP03LED GP04LEDFluorescent lightning Blue LED (470 nm) Blue/Green LED (470 - 520 nm)

Standard lightning White light LED (lid panel) White light LED (lid panel)

White light LED (Epi) White light LED (Epi)

Image format JPEG TIFF, BMP, JPEG

Sensor CMOS CMOS

Resolution 9 MPixel 9MPixel

Image storage Wi-Fi ready (PQI) SD-Card USB 2.0

Exposure setting 11 scales (-5 EV to 5 EV) 11 scales (-5 EV to 5 EV)

Focal length Fixed focal length Fixed focal length

View area 16 cm x 11.5 cm 16 cm x 11.5 cm

Filter Amber Filter (~520 nm) Amber Filter (~520 nm)

Display Build-in 2.7“ screen Build-in 2.7“ screen

Output TV-out (3.5 mm and RCA end) VGA, USB

Thermal printer ü Yes (analog) ü Yes (digital)

Power AC Adapter, 12 V / 1.5 A AC Adapter, 12 V / 4.16 A

Dimension 19.8 x 22.7 x 25.4 cm 23 x 23 x 21 cm

Accessories included Gel shovel, amber filter shield Gel shovel, amber filter shield

Weight 3.2 kg 3.5 kg

Overview

Imaging Systems

Cat. No. GP05LED GP-FAS-VFluorescent lightning Blue/Green LED (470 - 520 nm) Blue/Green LED (470 - 520 nm)

White light imaging* FastGene®White Light Plate Yes

Image format JPEG, RAW TIFF, BMP, JPEG, PNG

Sensor LIVE-CMOS CCD

Resolution 16 MPixel 2 MPixel (CCD), 12 Bit Grayscale

Image storage SD-Card internal Storage/ USB 2.0

Exposure setting 1/4000‘ to 30‘ 1/1000 to 30‘

Focal length (eq. 35mm) ~12 - 32 mm (128 mm digital zoom) 12.5 - 75 mm (f/1.2)

Focus TTL-automatic focus manual focus

View area 20 cm x 16 cm 26 cm x 21 cm

Filter Amber Filter (~520 nm) Amber Filter (~520 nm)

Display 3“ tilt- and touch-screen 10.4“ touchscreen

Output MicroHDMI, USB, SD-Card USB

Thermal printer Yes (analog) Yes (digital)

Power supply 100 - 220 V, 50 - 60 Hz 100-220 V, 50 - 60 Hz

Dimension 50 x 26 x 34 cm 38 x 40 x 78.5 cm

Accessories included Amber goggles Image capture and editing software

Weight 6.3 kg 35 kg

*In combination with the FastGene White Light Plate (Cat. No. FG-07)

FAS Digi FAS-V

Overview

FAS-V

Key Features• Very sensitive 2MPixel CCD-Camera (0.13 Lux)

• 10.4” Touchscreen display

• 26 cm x 21 cm Blue/Green LED Transilluminator

• 26 cm x 21 cm White LED Transilluminator

The FastGene® FAS-V is our most advanced imaging system. It is a standalone imaging system with an internal computer and a 10.4“ touchscreen. It contains our largest imaging area yet, for DNA as well as for protein imaging. The super sensitive CCD-Sensor is combined with a high-end parfocal lens, normally only found in microscope. The combination of our unique excitation technology with the high-end components deliver the best results with an easy to use system.

• Detection and documentation of DNA/RNA dyes

• Detection of GFP, eGFP, YFP, etc.

• Documentation of protein gels

• Documentation of Petri dishes & membranes

• On-side image editing

Applications

Image formats• Jpeg

• TIFF

• BMP

• PNG

Blue/Green LED TransilluminatorUnique Excitation Technology

• Enables the detection of red and green DNA dyes• Huge imaging area is 26 cm x 21 cm big • Superb uniform transillumination• 12 large LED arrays at each side

White LED Transilluminator• Detection of protein • Very large white LED table (26 cm x 21 cm)• Directly controlled by the touchscreen

White LED Room light• Documentation of membranes and Petri dishes• Two white LED arrays illuminate the imaging box

CCD-Sensor• CCD-Sensor size of 1/1.8“• Pixel size of 4.4 µm • Detection of very low signals of 0.13 Lux*• Images can be recorded in TIFF, JPEG, BMP and PNG format• 16 GB internal SSD storage or external USB-stick• Exposure time from 0.001 sec up to 30 sec• Manufactured in Japan

*Clarity of a normal day is between 20 000 and 100 000 Lux

Very bright parfocal lens - no more focusingThe lens of the FastGene® FAS-V is parfocal (preinstalled focus does not have to be readjusted when zooming in or out)• Huge aperture* of f/1.2• Aperture is regulated steplessly, • 6 x Zoom • Focal length of 12.5 mm to 75 mm

*opening of the lens iris

Optical Unit

Illumination

Comparison

The FAS V Imaging System has a highend Japanese CCD-sensor able to detect 0.13 Lux

2 MPixel UXGA CCD Sensor

10.4” Touchscreen Display

Intuitive Software

White LED room light

Blue/Green LED Transilluminator

White LED Transilluminator

The Blue/Green LED technology is able to excite red and green DNA dyes, e.g. MIDORIGreen, SYBR™ & EtBr

FAS-V

The imaging software takes 6 shots simultaneously, with different exposure times and includes an image editor

Ethidium Bromide MIDORIGreen Direct MIDORIGreen Advance

TouchscreenThe FastGene® FAS-V is controlled by a 10.4“ touchscreen

• All three light sources can be activated or deactivated by the touchscreen.

• Exposure time and gain adjustment can be adjusted using the slide function or by directly adding the desired value.

• Freezing the image

• Capturing the image

• Inverting live view or loaded image

Image EditingRecorded images can be directly edited

• Adjusting contrast

• Cropping the image

• Rotation of the image anti- and clockwise

• Mirroring vertically and horizontally

• Inverting black and white

ConnectionsStorage

The internal SSD-storage of the FastGene® FAS-V has a size of 16 GB. This can be expanded using an external USB-Stick. There are 3 USB ports to connect USB-sticks, mouse or a keyboard

NetworkThe FastGene® FAS-V can be directly connected to a network using the 100 MBits ethernet port

PrintingThe FastGene® FAS-V can be used with the thermal printer Mitsubishi P95D

FAS-V

Camera sensor type 2MPixel CCD - charge-coupled deviceSensor and image size 1/1.8” Sensor with 1600 x 1200 (UXGA)Mininmal illumination 0.13 LuxImage format TIFF, JPEG, BMP, PNGExposure time 0.001 to 30 secPixel size 4.4 x 4.4 µmAperture f / 1.2Focal distance 12.5 - 75 mm Zoom range 6 x zoom Filter Removable amber filter

Display 10.4” Color touchscreen Display resolution 800 x 600

Internal storage 16 GB SSDExternal storage USB 2.0Connections Ethernet (100Mb), USB 2.0, Thermoprinter Mitsubishi P95D supportedSoftware Touchscreen with image editor software

Light sources Blue/Green LED transilluminatorWhite LED transilluminatorWhite LED room light

Transilluminated area 26 x 21 cm (both)Fluorescent light 470-520 nmBlue/Green LEDs 24 x spots

Rated Voltage 100-240 V, 50 / 60 Hz, 2 A

Footprint (WxDxH) 38.2 cm x 40 cm x 78.5 cmWeight 35 Kg

Specification

FAS-V

FAS-Digi

Key Features• Very high resolution - 16 MPixel LiveMOS Sensor

• 3” tiltable touchscreen display

• 20 cm x 16 cm Blue/Green LED Transilluminator

• White LED Transilluminator compatible

• Controlable by smartphones and tablets

The FastGene® FAS Digi was the first imaging system to use the Blue/Green LED imaging technology, and was designed to be a modular system. It is composed by the FastGene® Blue/Green LED Transilluminator (FG-08) and a high-end camera. It has the highest resolution of all of our imaging system with incredible 16 MPixel. The camera is Wi-Fi enabled and can be controlled using a smart device.

• Detection of red and green DNA/RNA dyes




• Excision of DNA from agarose gels

Applications


• RAW


• Enables the detection of red and green DNA dyes

• Imaging area of 20 cm x 16 cm

• Superb uniform transillumination

• 6 large LED arrays at each side

Illumination

LiveMOS-Sensor• 24 mm - 64 mm focal length

(256 mm with digital zoom)

• 16 MPixel

• Focus by touchscreen

• Images can be recorded in JPEG and RAW format

Optical Unit

FAS Digi

16 MPixel LiveMOS Camera

Aluminum zoom ring

Amber filter

Metal dark box(removable)

Blue/Green LEDTransilluminator

3“ Touchscreen

Wi-Fi with tethering function

Highest resolution and ultra sharp pictures. Images can be printed in DIN A2 format


Comparison

B/G LED GelPicBox

Key Features• Smallest documentation system

• 2.7” LCD display

• 16 cm x 11.5 cm viewing area

• Blue/Green LED Transilluminator

• USB 2.0 port

The FastGene® Blue/Green LED GelPicBox combines a compact footprint with the advantages of the Blue/Green LED technology. This is the probably smallest imaging system in the world.The FastGene® Blue/Green LED GelPicBox is equipped with Blue/Green and white LEDs to document nucleic acids as well as protein gels and membranes.

• Detection of red and green DNA/RNA dyes




• Excision of DNA from agarose gels

Applications


• TIFF

• BMP


• Enables the detection of red and green DNA dyes

• Imaging area of 16 cm x 11 cm

• Superb uniform transillumination

• 12 LED arrays at each side

White LED Lid• Detection of protein

• White LED in the lid

White LED Room light• Documentation of membranes and Petri dishes

• Two white LED arrays illuminate the imaging box

Illumination

CMOS-Sensor• Fixed focal length

• 9 MPixel

• Images can be recorded in TIFF, JPEG and BMP format

• External USB-stick

Optical Unit

Blue/Green & White light selection switch

B/G LED GelPicBox

Upper cover lid

White backlight

LED white light

Power switch

Scan button

Menu button

USB slot

LCD display


Comparison

Illuminators

Cat. No. FG-08 FG-11Fluorescent lightning Blue/Green LED (470 - 520 nm) Blue/Green LED (470 - 520 nm)

DNA Dyes Compatibility Green and red dyes Green and red dyes

View area 20 cm x 16 cm n.a.

Filter Amber goggles (~520 nm) Amber filter (~520 nm) - 7 x 16 cm

Power AC adapter, 24 V / 1 A AC adapter, 24 V / 1 A

Dimension 19.8 x 22.7 x 25.4 cm 19 x 3.9 x 2.5 cm

Weight 2.9 kg 0.17 kg

Transilluminator Flashlight• Excitation of red and green dyes • Safe LED light

• No DNA degradation

• Better efficiency of downstream applications

• Safe working environment

• Long LED bulb life (50 000 h)

• Excitation of red and green dyes • Safe LED light

• No DNA degradation

• Mobile

• Real-time electrophoresis

• Stand available

Illuminators

Blue LED Transilluminator

FG-05 FG-06 FG-300Blue LED (470 nm) Blue LED (470 nm) UV-light (302 nm)

Green dyes Green dyes Red and green dyes

12 cm x 7 cm 20 x 16 cm 26 cm x 21 cm

Amber filter (~520 nm) Amber filter (~520 nm) UV-blocking shield

AC adapter 24 V, 1 A 24 V, 1.67 A 48 W

21 x 21 x 3 cm 38 x 40 x 8 cm 28 x 34 x 8 cm

2.1 kg 3 kg 4.3 kg

Blue LED Illuminator UV-Transilluminator• Safe LED light

• Compatible with green dyes

• Compact foot print

• Affordable

• Safe LED light

• Compatible with green dyes

• Large viewing area

• Powerful UV-filter

• High-quality UV-bulbs

• Low and high energy mode

• Large viewing area

Traditional Illuminators

Illuminators

NIPPON EUROPE

Binsfelder Straße 77D-52351 DürenGermany

[email protected]

+49 2421 55 496 0+49 2421 55 496 11

gel documentation systemsuv-light for up to 60 seconds. the pcr product was subsequently isolated...

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