Matt:LabNotes/2014-5-27

From ZhangLabWiki
Revision as of 00:27, 28 May 2014 by >Mzcai (Created page with "==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''' #Aspi...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

Artifical Rolonies

Procedure

  1. Add 140ul 100nM MALAT1_template to each well
    • Pre-heat to 85C
    • Incubate @RT for 30 min
      • C4 & C5 Incubate @RT for 15 min
  2. Aspirate & Wash once with 1X PBS
  3. 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