Matt:LabNotes/2014-5-27
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Artifical Rolonies
Procedure
- Add 140ul 100nM MALAT1_template to each well
- Pre-heat to 85C
- Incubate @RT for 30 min
- C4 & C5 Incubate @RT for 15 min
- Aspirate & Wash once with 1X PBS
- Capture with 10pM ppMALAT1
- Incubate @45C for 24 hours (5:30p- )
Component | Volume |
10X Ampligase Buffer | 80 |
1nM ppMALAT1 | 80 |
5 U/ul Ampligase | 80 |
H2O | 560 |
Total | 800 |
Exo I/III Test
Procedure
- Strip with 80% formamide for 15min @RT
- Wash with ddH2O twice
Experimental & Negative Control (Positions 3-12)
Component | Volume |
10X Ampligase Buffer | 50 |
2uM ppMALAT1 | 2.5 |
H2O | 447.5 |
Total | 500 |
Positive Control (Positions 1-2)
Component | Volume |
10X Ampligase Buffer | 10 |
H2O | 90 |
Total | 100 |