pcr gel

Tucson High SchoolBiotechnology Course

Spring 2010

What is gel electrophoresis?

- - -- - - -

++++++

wells DNA (-)

Loading Buffer

pcrproducts

+

STAIN

DNA Sequencing

DNA Sequencing: Basic Principle

Sequence of base pairs??

A ACGG AT CA TTCG

polymerase

first human genome = ten years @ $1 billion

The quest for the $1K human genome

2nd: 10,000 instrument days (30 instruments for 1 year) @ $10 million

J. Craig Venter3rd: 454 pyrosequencing, two months @ $2 million

James Watson

Nature, Nov 2008 -- “effectively” 4 human genomes (@$100K each)- first African genome (Illumina, UK)- first Asian genome (Beijing Genomics, China)- first cancer genome (Washington University, USA)

The $1,000 human genome, coming soon?

454 Pyrosequencing - the generations

The “pyro” in pyrosequencing

http://www.pyrosequencing.com/

Pyrosequencing - Library construction

Images courtesy of Roche (Technical presentation)

Pyrosequencing - emPCR

Pyrosequencing - sequencing

Pyrosequencing - image processing

Seq

uen

ce p

rod

uct

ion

(B

illi

on

s of

base

s/m

on

th)

Seq

uen

ce p

rod

uct

ion

(B

illi

on

s of

base

s/m

on

th)

0.500.50

0.800.80

1.00 1.00

0.300.30

0.400.40

0.200.20

0.100.10

00

0.700.70

0.600.60

0.900.90 Cost: C

en

ts per b

ase

Cost: C

en

ts per b

ase

.6.6

1.01.0

.8.8

.4.4

.2.2

00

1.61.6

1.41.4

1.21.2

2.02.0

1.81.8

3.13.1

19891989 19911991 19931993 19951995 19971997 19991999 20032003 2005200520012001

Courtesy of Phil Hugenholtz (JGI)

Sequencing costs