Matt:LabNotes/2016-4-11: Difference between revisions

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>Mzcai
(Created page with " #Wash with nf-H2O twice #Prepare Ampligase mix on ice #*Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme *Combine B11 and B12...")
 
>Mzcai
mNo edit summary
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#*Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme
#*Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme
*Combine B11 and B12: 100ul at ~200nM
*Combine B11 and B12: 100ul at ~200nM
**B12 looked to have some very fine PA gel still in there (not filtered enough?)
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Revision as of 01:38, 12 April 2016


  1. Wash with nf-H2O twice
  2. Prepare Ampligase mix on ice
    • Preheat H2O + Padlock probes to 85C and then snap cool before adding Ampligase buffer and enzyme
  • Combine B11 and B12: 100ul at ~200nM
    • B12 looked to have some very fine PA gel still in there (not filtered enough?)
Component Volume
DEPC-H2O 47
Ampligase Buffer 10
Batch 11 & 12 Padlock Probes ~200nM]] 33
Ampligase 10
Total 100
  1. Add mix to sample and incubate for 30min at 37C
  2. Move sample to 60C and oven slowly decreases to 55C and held for ~24hr