Rui:LabNotes/Haplotyping/2011-4-20

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Cell lysis[edit]

  • Alan gave me 1ml lymphocytes containing 1.23X10(6) cells
  • Spin down at 300g for 3min at R.T -- a clearly tight pellet
  • Wash with PBS and spin down at 500g for 3min at 4C -- a clear tight pellet
  • Add 200ul Kun's lysis buffer and pippet up and down by following his note on 8/19/2008 [1]-- immediately form a stretched aggregation of DNA/Ptn (?)
* In Kun's lysis buffer, it seems to contain the proteinase which directly remove Proteins and release gDNA
* Spin down and EtOH precipitation to check DNA in both sup. and pellet
* Probably, I should resuspend cells in TE buffer and then dilute to 1000cells in lysis buffer and quickly aliquot -- with 1000 cells, aggregation is probably not easy to form before aliquot.