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Old 12-02-2010, 01:05 PM   #27
Location: Connecticut

Join Date: May 2010
Posts: 42

Has anyone tried the method that Phillip suggests for avoiding the concatamer issue with the SMART kit? I was wondering if people thought is was successful and worth doing. I was unsure about how common a problem this was. Is this due to the enzyme specifically? Can it actually add the C's to a DNA template? I was a little surprised to see some many people with a similar issue as we use the Clontech kit for cDNA synthesis in the our lab regularly for other applications and haven't noticed these type of problems. Is this issue just due to long RT reaction times? Also, did downstream amplification of first-strand products work fine? Wouldn't this contribute to some 3' bias in sequencing? Just curious what peoples thoughts were.


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