There are peaks from 70 – 90 bp in addition to the main library peak on the Agilent™ 2100 Bioanalyzer chart?

Peaks from 70 – 90 bp (see the graph below) are primer dimers from the 2nd PCR and result from incomplete removal of small molecular materials during the final magnetic bead purification. These are usually caused by inaccurate pipetting of magnetic beads when making a large number of libraries in a short period of time OR insufficient removal of ethanol during washing steps. To remove any possible small-molecular-weight materials, it is wise to pool all indexed libraries (that will be sequenced in the same lane) and do one additional round of 1.2X magnetic bead purification, before the quantification step prior to sequencing.

2018-03-27T15:29:53+00:00 December 28th, 2016|Comments Off on There are peaks from 70 – 90 bp in addition to the main library peak on the Agilent™ 2100 Bioanalyzer chart?