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Old 11-02-2009, 09:07 AM   #11
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Location: France

Join Date: Nov 2009
Posts: 2
Default Does a broken T primer help ?

Hello everybody
We had on our first cDNA sequencing try, using a MINT kit used on a 1/4 Titanium plate. No concatenates, but a bimodal length distribution with a first peak around 70 nt and a second around 500 nt; about 50000 reads are below 100 nt, and 100000 above. Reads with a 5' polyT make up 50 % of the short, bad class. We used a first strand primer with 30 T, which I suspect to bring difficulties with pyrosequencing. Does anybody has experience with broken T primers ?

Thanks ahead!
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