Daniel:Notebook/HiResChrPaint/2014-9-10

From ZhangLabWiki
Revision as of 17:52, 10 September 2014 by >Djacobse (→‎DpnII/USER Digestion)
Jump to navigation Jump to search

Padlock Probes II

Back to Calendar

Nanodrop Results

DpnII/USER Digestion

Reaction Table

Reagent PPRL (uL)
amplicon 30
ug Input 2.0
USER 5
nf H2O 45
Total 80
  1. Set up reaction according to table in a 0.2 mL PCR tube
  2. Incubate for 2 hours at 37C
  3. Add 15 uL according to DpnII Primer table (below)

DpnII Primer Table

Reagent PPRL (uL)
10X DpnII Buffer 10
100 uM RE-DpnII-V4 guide oligo 5
Total 15
  1. Incubate at 94C for 2 minutes
  2. Cool to 37C and incubate for 3 minutes
  3. Add 5 uL DpnII and incubate at 37C for 2 hours
  4. Heat inactivate DpnII at 65C for 20 minutes

Nanodrop Results

Sample ng/uL ssDNA ug in 20 uL Total
PPRL-DpnII-A 38.7 0.8
PPRL-DpnII-B 35.3 0.7 1.5