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Elektrophorese-Technik

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DNA Electrophoresis

Dez 17, 2008

In all DNA-electrophopresis methods the DNA run negatively charged to the anode!
There are 3 different main DNA-electrophoresis methods:

Native samples run in native gels: d(ouble) s(dranded)-DNA electrophorese
ds: Easy method, 1 fragment = 1 band, but no exact molecular weights
(because of the double-stranded conformation polymorphisme)

Denaturated samples run in denaturated gels: s(ingle)-s(tranded) denatured electrophoresis
ss: Difficult method, 1 fragment = 2 bands (PCR-anomaly), but exact molecular weights
(method used in all sequencers)

Denaturated samples run in native gels: s(ingle)-s(tranded) conformation polymorphisme electrophoresis
sscp: No molecular weights but sequence-related run (= mutation-detection)

1st step:
Your target molecules --> which method, gel-matrix, buffer?

Range

Method

Gel

Buffer

Res

AN

 

Lenghts-Polymorphisme:

 

 

 

 

Narrow: 10 - 40 bp

15%T, native - semidisc (ds)

1001-24

1002-19

2bp

1003

Narrow: 10 - 100 b

15%T, denaturing - semidisc (ss)

1001-24

1002-19

1 b

1003

Narrow: 50 - 150 b

15%T, denaturing (ss)

1001-24

1002-19

2b

1012

Narrow: 120 - 250 b

10%T, denaturing (ss)

1001-02

1002-19

2b

1012

Narrow: 40 - 200 bp

HyRes, native (ds)

1001-25

1002-21

2 bp

1015

Narrow: 200 - 400 b

10%T, denaturing

1001-02

1002-19

2 b

1027

Broad:  50 - 2000 bp

10%T, native (ds)

1001-02

1002-08

8 bp

1018

Broad:  20 - 1500 bp

15%T, native (ds)

1001-24

1002-08

6 bp

1018

 

Mutation-Detection:

 

 

 

 

no molweights

Sequence-related run: (sscp)

1001-05
1001-24

1002-08
1002-19
1002-23

99%M

AN 1011

2nd step:
Select your slot-geometry ---> 25S - 36S - 52S - 103S