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

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'''Reaction Table'''
'''Reaction Table'''
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
{| 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"
|- style="background-color:#B8CCE4;font-size:12pt;font-weight:bold" align="center"
| width="93" height="33" | Reagent
| width="65" height="30" | Reagent
| width="65" | PPRL (uL)
| width="71" | PPRL-A1 (uL)
| width="71" | PPRL-A2 (uL)
| width="71" | PPRL-B1 (uL)
| width="65" | PPRL-B2 (uL)


|- style="font-size:12pt"
|- style="font-size:12pt"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | amplicon
|style="background-color:#FCD5B4" height="15"  valign="bottom" | amplicon
| align="center" align="center" valign="bottom" | 30
| align="center" align="center" valign="bottom" | 50
| align="center" align="center" valign="bottom" | 50
| align="center" align="center" valign="bottom" | 50
| align="center" align="center" valign="bottom" | 50


|- style="font-size:12pt"
|- style="font-size:12pt"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | ug Input
|style="background-color:#FCD5B4" height="15"  valign="bottom" | ug Input
| align="center" align="center" valign="bottom" | 2.0
| align="center" align="center" valign="bottom" | 94.2
| align="center" align="center" valign="bottom" | 94.2
| align="center" align="center" valign="bottom" | 103.4
| align="center" align="center" valign="bottom" | 103.4


|- style="font-size:12pt"
|- style="font-size:12pt"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | USER
|style="background-color:#FCD5B4" height="15"  valign="bottom" | USER
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 5


|- style="font-size:12pt"
|- style="font-size:12pt"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | nf H2O
|style="background-color:#FCD5B4" height="15"  valign="bottom" | nf H2O
| align="center" align="center" valign="bottom" | 45
| align="center" align="center" valign="bottom" | 30
| align="center" align="center" valign="bottom" | 30
| align="center" align="center" valign="bottom" | 30
| align="center" align="center" valign="bottom" | 30


|- style="font-size:12pt;font-weight:bold"
|- style="font-size:12pt;font-weight:bold"
|style="background-color:#FCD5B4" height="15"  valign="bottom" | Total
|style="background-color:#FCD5B4" height="15"  valign="bottom" | Total
| align="center" align="center" valign="bottom" | 80
| align="center" align="center" valign="bottom" | 85
| align="center" align="center" valign="bottom" | 85
| align="center" align="center" valign="bottom" | 85
| align="center" align="center" valign="bottom" | 85


|}
|}

Revision as of 18:30, 10 September 2014

Padlock Probes II

Back to Calendar

Nanodrop Results

Sample 1:1 ng/uL 5:1 ng/uL 5:1 ng/uL Projected 10:1 ng/uL 10:1 ng/uL Projected Average ng/uL Standard Deviation ug in 100 uL
PPRL-A 1944.1 373 1865 184.4 1844 1884.4 52.8 188.4
PPRL-B 2064.8 421.7 2108.5 203 2030 2067.8 39.3 206.8

Usually after ethanol precipitation I get really high results, so I'll just say these are fine and continue as though I have a reasonable amount of material.

DpnII/USER Digestion

Reaction Table

Reagent PPRL-A1 (uL) PPRL-A2 (uL) PPRL-B1 (uL) PPRL-B2 (uL)
amplicon 50 50 50 50
ug Input 94.2 94.2 103.4 103.4
USER 5 5 5 5
nf H2O 30 30 30 30
Total 85 85 85 85
  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