Kun:LabNotes/ASE/2007-12-14

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Exp.1 PCR on the circularized CES22k probes[edit]

                              one reaction       12 reactions
          Template:             10ul                  each
          10x Buffer:            9ul                 108ul
          25mM MgCl2           5.4ul                64.8ul
          25mM dNTP            1.2ul                13.2ul
          100uM AmpSolV6F2     0.4ul                 4.8ul
          100uM AmpSolV6R2     0.4ul                 4.8ul
          JumpStart Taq          1ul                  12ul
          50X SYBG I           0.8ul                 9.6ul
          H2O                   72ul                 861ul

94C 3min -> 8 cycles of (94C 45sec -> 57C 1min -> 72C 1min) -> 10 cycles of (94C 45 sec -> 72 1min) -> 72C 3min -> 15C hold.

Took out 1C and 2A, add 4 more cycles on the remaining reactions.

  File:2007-12-17-CES22k-SolLib-PAGE-quant.png
    [12/15-12/17] PAGE purification:
    NA20431    2.48ng/ul     28nM
    GM20431    3.15ng/ul     36nM
    GC1F       2.05ng/ul     23nM
    GC1EP      0.58ng/ul    6.6nM
    Hues6g     2.69ng/ul     31nM
    Hues6c     2.53ng/ul     29nM
    H1g        3.09ng/ul     35nM
    H1c        1.41ng/ul     16nM
    H2g        4.08ng/ul     46nM
    H2c        0.68ng/ul    7.7nM
    Repeat the PAGE quantification on 12/19/2007
    H2g        5.48ng/ul 
    H2c        0.46ng/ul
    BJc        2.08ng/ul


I got 118,945 reads from NA20431 in the Dec-19 run. The correlation of read number with probe abundance is R=0.145.