Non-radioactive DNA sequencing

Details Non-radioactive DNA sequencing Non-radioactive DNA sequencing

1994-03-23

1996-06-04

Jones, W. Gary

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

204462, C12Q 168, G01N 2726

Patent

active

055232060

ABSTRACT:
A new process and kit are described that combines methods for generating the nucleotide base sequence of a DNA molecule with an ultra-sensitive silver staining protocol. This new combination of technologies allows for a direct, non-instrument based visualization of electrophoretically separated sequencing fragments. This non-radioactive system includes sequencing the DNA molecule by forming a set of fragments using an enzymatic dideoxy-mediated chain termination method, electrophoretically separating the DNA fragments on a gel medium, and exposing the gel medium to ultra-sensitive silver-staining solutions for a time determined by viewing the silver stain reacted primer extension products.

REFERENCES:
Beidler et al., Anal. Biochem 126(2):374-380 (1982) "Ultra sensitive stainring of nucleic-acids with silver . . . ".
Bassam et al., Anal. Biochem 196:80-83 (1991) "Fast and sensitive silver staining of DNA . . . ".
Maxam et al., P.N.A.S. 74:560-564 (1977) "A new method for sequencing DNA".
Sanger et al., P.N.A.S. 74:5463-5467 (1977) "DNA Sequencing with chain-terminating . . . ".

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