FLX vs Titanium library

[krelmer]

Hello,

Last year we had a normalized cDNA library made by MWG and sequenced on 454 FLX with standard chemistry. One month later Titanium chemistry was released and we then learned that our very expensive normalized libraries could only be used with standard chemistry and never with Titanium. MWG should have told us this before hand!!! Anyway, I realise that the libraries fundamentally differ, but I am wondering if anyone knows of a homemade method to upgrade our libraries? Even if we lose some read length it would still be better than reverting to an earlier chemistry (we get 530 Mb with cDNA in Titanium and only got 115 Mb with standard chemistry).

Thanks!

Kathryn

[flxlex]

In theory, the only way I could think of converting the library would be by putting Titanium adaptors on the molecules. However, the library is single stranded, meaning it would have to be converted to dsDNA first. Also, the amount of library material is relatively minute. Perhaps amplification could help here, but that is going to introduce some bias.

So in practice, I am afraid you are out of luck...

[krelmer]

Ok, thanks for the tip!

[nextgen]

Can you design a new PCR primer to include part of old adaptor sequence and the new adaptor sequence? When you re-amplify your library for a few cycles, the new adaptor sequence will be added.