I'm having a strange problem with my library prep for Illumina. Unfortunately I have no been able to find any information on this, not here nor from Agilent, so i have to assume that i'm the only one with this problem...
maybe i can get some feedback from you guys
My post-PCR RAD libraries (i.e. dsDNA 300-500bp) are clearly visible on my agarose gels
but hardly or mostly not at all on the DNA1000 Bioanalyzer chips
The Bioanalyzer runs look fine, i.e. the ladder is excellent and I cannot detect any problems with the lower and higher marker peaks, i.e. the matrix and peak detection seems to work fine. Between those peaks though the electrogram is completely flat, no DNA at all by now I have at least 20 different PCR products that I can't visualize on the BioAnalyzer. So my PCR apparently works but without the Bioanalyzer I can't sequence
Has anybody seen/heard of this before? any idea what I can do?? I guess the problem is most likely with the Bioanalyzer but...
any comments appreciated!!
cheers
P
maybe i can get some feedback from you guys
My post-PCR RAD libraries (i.e. dsDNA 300-500bp) are clearly visible on my agarose gels
but hardly or mostly not at all on the DNA1000 Bioanalyzer chips
The Bioanalyzer runs look fine, i.e. the ladder is excellent and I cannot detect any problems with the lower and higher marker peaks, i.e. the matrix and peak detection seems to work fine. Between those peaks though the electrogram is completely flat, no DNA at all by now I have at least 20 different PCR products that I can't visualize on the BioAnalyzer. So my PCR apparently works but without the Bioanalyzer I can't sequence
Has anybody seen/heard of this before? any idea what I can do?? I guess the problem is most likely with the Bioanalyzer but...
any comments appreciated!!
cheers
P
Comment