Hi guys!
I keep having a problem with the final RNA-seq liabrary size. After enrich DNA fragments, everything seems to be OK on the electrophoretogram with DNAmaker I (see attached thumbnails) and we chose gel-method to obtain the fragments (300-400bp) and purified using MinElute Gel Extraction Kit (QIAGEN). BTW, we eluted the liabrary by 25ul RNA-free water instead of QIAGEN buffer EB.
Then we used Agilent Bioanalyzer to obtain the exact fragments size and found every sample had a unexpected signal between 700-1000bp region!(see attached thumbnails)
Does anyone have this problem before? Do you have any idea what can cause this unexpected signal? I swear no irrelevant fragments were extracted during my experiments.
Many thanks!!!
I keep having a problem with the final RNA-seq liabrary size. After enrich DNA fragments, everything seems to be OK on the electrophoretogram with DNAmaker I (see attached thumbnails) and we chose gel-method to obtain the fragments (300-400bp) and purified using MinElute Gel Extraction Kit (QIAGEN). BTW, we eluted the liabrary by 25ul RNA-free water instead of QIAGEN buffer EB.
Then we used Agilent Bioanalyzer to obtain the exact fragments size and found every sample had a unexpected signal between 700-1000bp region!(see attached thumbnails)
Does anyone have this problem before? Do you have any idea what can cause this unexpected signal? I swear no irrelevant fragments were extracted during my experiments.
Many thanks!!!
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