Matt:LabNotes/2014-2-6: Difference between revisions

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>Mzcai
(Created page with "===ppMALAT1 Capture in vitro=== *From yesterday Matt:LabNotes/2014-2-5 will want to do atleast 10x the amount Alan did AlanFung:LabNotes/Capturing/2014-1-23 *Will do ...")
 
>Mzcai
Line 22: Line 22:
  -> add 3ul ampligase mix (0.5U/ul AmpLigase)
  -> add 3ul ampligase mix (0.5U/ul AmpLigase)
  -> 55C 5h-> 94C 2min -> add 2ul Exo I/III mix-> 37C 2h -> 94C 2min -> 4C hold.
  -> 55C 5h-> 94C 2min -> add 2ul Exo I/III mix-> 37C 2h -> 94C 2min -> 4C hold.
*Stored in -20C until purification

Revision as of 01:30, 8 February 2014

ppMALAT1 Capture in vitro

  • Will do 8 reactions of 35ul, so almost 19x more
' 100uM MALAT1 Template H2O 200uM ppMALAT1 10X Ampligase Total
1 Reaction 6 18 3 3 30
8 Reactions 48 144 24 24 240
  • Chose to use more template and less padlock probe than Alan because we want all padlock probes to circularize
    • So more like 12x more but hopefully with higher yield
Program stored under Kun/CPG
-> 95c 30sec -> cool down to 55C at 0.02C/sec -> 55C 16.5h (started at 6pm)
-> add 3ul ampligase mix (0.5U/ul AmpLigase)
-> 55C 5h-> 94C 2min -> add 2ul Exo I/III mix-> 37C 2h -> 94C 2min -> 4C hold.
  • Stored in -20C until purification