How to Make Nucleotide Ladder Markers?

Bert Gold Bert_member at newsguy.com
Fri Nov 26 19:13:36 EST 2004


After Duncan posted his reply, I found:
DeWoody JA. Schupp J. Kenefic L. Busch J. Murfitt L. Keim P. Universal
method for producing ROX-labeled size standards suitable for automated
genotyping. [Article] /Biotechniques. 37(3):348-+, 2004 Sep.

and

http://life.biology.mcmaster.ca/~brian/evoldir/Other/DNA.ladder.answers

I'm still not satisfied that I've gotten to the best
approach, however.

I am thinking about some ligase based method.

Anyhow, I am still open to ideas.

In article <rszESSAbewpBFAAl at abuse.plus.com>, Duncan Clark says...
>
>Historians believe that in newspost <co57q301hgj at drn.newsguy.com> on 
>Thu, 25 Nov 2004, Bert Gold <Bert_member at newsguy.com> penned the 
>following literary masterpiece:
>>How would you make a ladder that
>>consists of: 60, 70, 80, 90, 100,
>>120, 140, 160, 180, 190, 200, 220,
>>240, 260, 280, 300, 320, 340, 360,
>>380, 400, 420, 440, 460, 480, 500,
>>520, 540, 560, 580, 600, 620, and
>>640 nucleotides?
>
>Umpteen individual PCRs - watch out for the +A addition?
>
>Or mutate or gene synthesise a DNA that has say C only at those 
>positions. Use a single fluorescent ddGTP sequencing reaction so only 
>those positions light up?
>
>In the latter case you can't use a fluorescent primer as you will 
>generate a fluorescent run-off product much much brighter than the 
>fragments you require.
>
>Duncan
>




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