Hosuk:LabNotes/2014-4-10

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Revision as of 21:51, 10 April 2014 by >Hosuki78
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Cancer Cell : Mutation detection test - 2nd Try

  • NCIH-1975 in 24 well plate
  • Fix : 04/08
  • Finish all process 04/10


  • Well 1, 2 : EGFR, BRAF RT Primers
  • Well 3, 4 : EGFR, BRAF RT Primers + BRAF Stop primer 200bp
  • Well 5, 6 : EGFR, BRAF RT Primers + BRAF Stop primer 200bp
  • Well 7, 8, 9, 10 : random hexamer


File:24Well 2014-04-10.png File:NCIH1975 2014-04-10 1stRolony Count.png


Result for 1st Rolonies

  • Well 1 ~ Well 6 have similar number of rolonies,
  • which the numbers are much less than Well 7, 8, and 9.
    • (Well 10 has few, it’s because of mistake at RNaseH step)


PGP1F sample for 13k Padlock probe test

  • Fix : 04/06, 2 dishes
  • Finish all process 04/10, imaging


  • S1 : incubate RCA primer at RT for 10min after incubating RCA primer at 60C oven for 15min.
  • S2 : Wash and add RCA mix right after incubating RCA primer at 60C oven for 15min.


Result for 1st Rolonies

  • Both S1 (incubate RCA primer at RT for 10min), S2 looked similar amount of Rolonies
  • Both sample have pretty good 1st Rolonies!
  • I will use S2 for Ampligase and RCA with13k Padlock probes.


File:PGP1F 2014-04-10 1stRolony Count.png