Daniel:Notebook/HiResChrPaint/2014-9-10: Difference between revisions

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>Djacobse
>Djacobse
Line 38: Line 38:
#Set up reaction according to table in a 0.2 mL PCR tube
#Set up reaction according to table in a 0.2 mL PCR tube
#Incubate for 2 hours at 37C
#Incubate for 2 hours at 37C
#Add 15 uL according to DpnII Primer table (below)
#Add 10 uL DpnII buffer (10X)
 
'''DpnII Primer Table'''
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="background-color:#B8CCE4;font-size:12pt;font-weight:bold" align="center"
| width="93" height="31" | Reagent
| width="65" | PPRL (uL)
 
|- style="font-size:12pt"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | 10X DpnII Buffer
| align="center" align="center" valign="bottom" | 10
 
|- style="font-size:12pt"
|style="background-color:#FCD5B4" height="30"  valign="bottom" | 100 uM RE-DpnII-V4 guide oligo
| align="center" align="center" valign="bottom" | 5
 
|- style="font-size:12pt;font-weight:bold"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | Total
| align="center" align="center" valign="bottom" | 15
 
|}
 
#Incubate at 94C for 2 minutes
#Incubate at 94C for 2 minutes
#Cool to 37C and incubate for 3 minutes
#Cool to 37C and incubate for 3 minutes

Revision as of 17:56, 10 September 2014

Padlock Probes II

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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 10 uL DpnII buffer (10X)
  4. Incubate at 94C for 2 minutes
  5. Cool to 37C and incubate for 3 minutes
  6. Add 5 uL DpnII and incubate at 37C for 2 hours
  7. Heat inactivate DpnII at 65C for 20 minutes

Nanodrop Results