strong 140bp peak following RNA library construction
Hello all,
I have created an illumina truseq library on bacteria by first performing ribozero rRNA depletion kit and then performing a truseq stranded mRNA prep while skipping the poly-A selection reaction. my libraries are complete and in cDNA format. i have run a DNA 1000 bioanalyzer assay to visualize the libraries. we see the desired ~260 bp library peak but in every sample there is also a strong ~140bp sharp peak which I think will throw off my molarity calculations (what i'm actually thinking about is proper flow cell cluster density), if it is say, a product of the library prep which cannot form clusters. in certain circumstances it seems to be in much higher concentration than the fragments that i want to sequence. does anyone know what this ~140bp sharp peak is in every sample and i'm also interested to know if there is some way of accounting for this when i do my normalization dilutions pre-pooling. thank you
I have created an illumina truseq library on bacteria by first performing ribozero rRNA depletion kit and then performing a truseq stranded mRNA prep while skipping the poly-A selection reaction. my libraries are complete and in cDNA format. i have run a DNA 1000 bioanalyzer assay to visualize the libraries. we see the desired ~260 bp library peak but in every sample there is also a strong ~140bp sharp peak which I think will throw off my molarity calculations (what i'm actually thinking about is proper flow cell cluster density), if it is say, a product of the library prep which cannot form clusters. in certain circumstances it seems to be in much higher concentration than the fragments that i want to sequence. does anyone know what this ~140bp sharp peak is in every sample and i'm also interested to know if there is some way of accounting for this when i do my normalization dilutions pre-pooling. thank you
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