Question: Mixed population of cells - Targeted / Amplicon Sequencing?
gravatar for alexille5640
4 months ago by
alexille564010 wrote:

I have the following:

  • SAMPLE 1: a culture of normal unmodified HEK293 cells (a pure population where all cells are the same)
  • SAMPLE 2: a culture of HEK293 cells on which I did a custom gene knock in via CRISPR and homology directed repair. (after a 90%+ transfection rate, only about 3% of cells sustained a stable knock-in as indicated by the GFP sequence i stuck on the end of my gene (which was designed to be knocked-in along with the gene))

I was told I could submit both samples to be sequenced and that NGS targeted sequencing / amplicon sequencing would be able to confirm whether or not my donor gene was inserted at my desired target sites. Is this correct?

(considering that sample 2 is a mixed population and only about 3% exhibit what appears to be a stable knock-in)

sequencing next-gen gene • 128 views
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