Daniel:Notebook/HiResChrPaint/2014-9-10: Difference between revisions
Jump to navigation
Jump to search
>Djacobse |
>Djacobse |
||
Line 47: | Line 47: | ||
'''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=" | | 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" | | | 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. | | 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" | | | 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" | | | 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
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 |
- Set up reaction according to table in a 0.2 mL PCR tube
- Incubate for 2 hours at 37C
- Add 10 uL DpnII buffer (10X)
- Incubate at 94C for 2 minutes
- Cool to 37C and incubate for 3 minutes
- Add 5 uL DpnII and incubate at 37C for 2 hours
- Heat inactivate DpnII at 65C for 20 minutes