Matt:LabNotes/2014-2-6
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ppMALAT1 Capture in vitro[edit]
- 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 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 in 1X Buffer) -> 55C 5h-> 94C 2min -> add 2ul Exo I/III mix-> 37C 2h -> 94C 2min -> 4C hold.
- Stored in -20C until purification