6.802/6.874/20.390/20.490/HST.506 Lecture April 2 2020 - Shared screen with speaker view
Seung-Hyun B Ko
Is barcode addition sequential?
how can the barcode enter the cell and anneal to every mRNA?
Is there a way to put UMI in the pool approach?
How does the template switch oligo limit the number of copies made?
@Meghana which technology was your question referencing?
I was referring to the method described to deal with PCR bias where you limit the number of times you can amplify a template sequence?
is multiplexing the same as sequencing different samples individually
is this cheaper?
@Meghana Using the template switch oligo is what enables the reverse transcription product to receive a unique molecular identifier (UMI). This UMI will enable you to determine if a PCR product was over amplified in your sequencing library. Because you have UMIs on each read, you can adjust the transcript abundance to be independent of the library abundance and better reflect the abundance in the cells.
That makes sense! Thank you!
What % of RNA is actually captured on individual beads that go on to be amplified and sequenced?