Amplified Library Prep Quality Control

Q: What quality control should I do after preparing amplified libraries?

A: Make sure to perform the following library quality control steps:

  1. Determine the concentration (ng/µL) of each library using the Qubit 1X dsDNA HS kit.
  2. After library purification, run the library on a Bioanalyzer, TapeStation, or equivalent. Load appropriate (ng) amount of library per recommended kit protocol to be within the acceptable detection range. When checking the quality of the library: (1) Visualize the size distribution to determine average fragment size (bp). (2) Ensure there are no other peaks of higher or lower molecular weight, other than the expected library and the upper and lower markers.

Note that Singular Genomics (SG) non-indexed primers are 62 bp long, while SG indexed primers are 75 bp long. To calculate the expected library size, take the expected insert size and add 124 bp for non-indexed runs or 150 bp for indexed runs.

Version: 29 Sep 2023