Kun:LabNotes/Haplotyping/2009-1-29: Difference between revisions

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(New page: ==Perform 2nd round MDA on the amplicons from 01/28/2009== *The purpose of this experiment is to generate more DNA for downstream analyses. Only the 16 amplicons from the haploid templates...)
 
mNo edit summary
Line 10: Line 10:
   RepliPhi Phi-29 (100U/ul)      10ul      30
   RepliPhi Phi-29 (100U/ul)      10ul      30
   Add 200ul MDA mix to each of the tube on cool block, 30C 2h, 85C 3min.
   Add 200ul MDA mix to each of the tube on cool block, 30C 2h, 85C 3min.
  Make aliquots of amplicons (50ul each) for Infinium Genotyping:
  GM18508A
  GM18507B
  GM18507D
  GM18507G
  GM20431A
  GM20431B
  GM20431C
  GM20431F
  Ethanol precipitation of 150ul amplicons:
  GM18508A
  GM18507B
  GM18507D
  GM18507G
  GM20431A
  GM20431B
  GM20431C
  GM20431G

Revision as of 22:23, 29 January 2009

Perform 2nd round MDA on the amplicons from 01/28/2009

  • The purpose of this experiment is to generate more DNA for downstream analyses. Only the 16 amplicons from the haploid templates will be re-amplified.
    Set up master mix:
                                         x 16
  10x RepliPhi phi-29 buffer     20ul     320
  1uM N6 primer                  10ul     160
  25mM dNTP                      8ul      128
  2X SYBR Green I                10ul     160
  H2O                           142ul    2272
  RepliPhi Phi-29 (100U/ul)      10ul      30
  Add 200ul MDA mix to each of the tube on cool block, 30C 2h, 85C 3min.
  Make aliquots of amplicons (50ul each) for Infinium Genotyping:
  GM18508A
  GM18507B
  GM18507D
  GM18507G
  GM20431A
  GM20431B
  GM20431C
  GM20431F

  Ethanol precipitation of 150ul amplicons:
  GM18508A
  GM18507B
  GM18507D
  GM18507G
  GM20431A
  GM20431B
  GM20431C
  GM20431G