Hi,
I have trying to combine the NEBNext sample prep kit with Sigma ordered Paired-end primers and adapters to prepare paired-ended libraries of genomic DNA....but i am having problems.
I recently made libraries with genomic DNA and observed a strange band at 120-150bp..this isn't primer/dimers as they run much lower nor the library. I do not do a size selection before the enrichment but after.
I tried to TA clone the band and observed lots of positive colonies and few negative ones. However, when i look at the results only a 5bp product has ligated (very weird).
Any ideas would be much appreciated.
Thanks
I have trying to combine the NEBNext sample prep kit with Sigma ordered Paired-end primers and adapters to prepare paired-ended libraries of genomic DNA....but i am having problems.
I recently made libraries with genomic DNA and observed a strange band at 120-150bp..this isn't primer/dimers as they run much lower nor the library. I do not do a size selection before the enrichment but after.
I tried to TA clone the band and observed lots of positive colonies and few negative ones. However, when i look at the results only a 5bp product has ligated (very weird).
Any ideas would be much appreciated.
Thanks
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