I am trying to making Illumina sequencing library using transposon based method to add adaptor. However, instead of using the adaptor used in Nexera kits, I would like to use the Y-shaped adaptor. Thus, the gap between the bottom stand of adaptor and bottom strand of genomic DNA need to be repaired. I want to find a method to do gap filling, however, I do not want the DNA to the 3' of the gap (i.e., bottom strand of the adaptor) to be removed by 5'-3' exonuclease activity of DNA Pol I and then re-synthesized. Can I use Klenow fragment here? It is also strange that no one seems to use a Y-shaped adaptor in transposon based library prep. Is there any technical difficulties here?
-
CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).
Despite this, “CRISPR helped turn genome editing from a specialized technique into...07-31-2026, 11:01 AM
Comment