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Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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DNA Electrophoresis

Mai 30, 2018

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)

[ds-DNA]
[ss-DNA]
[sscp-DNA]
[DNA-Trouble]

Range

Method

Res

 

Lenghts-Polymorphisme:

 

Narrow:  10  -  40 bp

15%T, native - semidisc (ds)

2bp

Narrow:  10  -  100 b

15%T, denaturing - semidisc (ss)

1 b

Narrow:  50  -  150 b

15%T, denaturing (ss)

2b

Narrow:  120  -  250 b

10%T, denaturing (ss)

2b

Narrow:  40 -  200 bp

HyRes, native (ds)

2 bp

Narrow: 200 -  400 b

10%T, denaturing

2 b

Broad:    50 - 2000 bp

10%T, native (ds)

8 bp

Broad:    20 - 1500 bp

15%T, native (ds)

6 bp

 

Mutation-Detection:

 

no molweights

Sequence-related run: (sscp)

99%M