Kenneth Day

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Questions related from Kenneth Day

For an in vitro enhancer screen assay, would you use linear construct design or plasmid based, STARR-Seq method?

Looking for anyone with experience in assessment of differences between linear construct design versus plasmid based, STARR-Seq method... https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4690393/?

08 August 2016 1,723 1 View

Does anyone have experience with immunoprecipitation or Dynabead pulldown of large TdT end labeled(biotinylated) restriction fragments (>10kb)?

Genomic DNA is digested with a restriction enzyme, end labeled with biotin-dNTP with TdT, and then immunoprecipitated with Protein G Dynabeads coated with anti-biotin antibody.

12 December 2014 9,980 15 View

For preparation of next-generation sequencing libraries, why is end repair performed at 20 degrees?

The subsequent step with Klenow exo- is done at 37 degrees to adenylate the blunt ended fragments. End repair uses a cocktail of T4 DNA polymerase, Klenow, and polynucleotide kinase.

12 December 2013 7,213 7 View