Dear All,
In the aim of a prokaryotic RNA sequencing on Roche 454; I used the kit Rapid cDNA Library (GS FLX Titanium).
To recover the small RNA, I modified slightly the protocol at the level of purification (calibration of beads Ampur) between the enzymatic steps (RT, Fragment repair, ..)
Unfortunately, it seems that the step of the ligation of double-stranded adapters doesn't work very well because the controls checked using Bioanalyser of Agilent (high sensitivity) show the presence of my product, but no matches were exploitable using the TBS fluorometer and at the stage of titration.
Has anyone already meet this kind of problem on the kit Rapid library?
thanks for your advices.
cheers.
In the aim of a prokaryotic RNA sequencing on Roche 454; I used the kit Rapid cDNA Library (GS FLX Titanium).
To recover the small RNA, I modified slightly the protocol at the level of purification (calibration of beads Ampur) between the enzymatic steps (RT, Fragment repair, ..)
Unfortunately, it seems that the step of the ligation of double-stranded adapters doesn't work very well because the controls checked using Bioanalyser of Agilent (high sensitivity) show the presence of my product, but no matches were exploitable using the TBS fluorometer and at the stage of titration.
Has anyone already meet this kind of problem on the kit Rapid library?
thanks for your advices.
cheers.
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