Daniel:Notebook/ComboLock/2017-3-21: Difference between revisions
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{| 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:#8DB4E2;font-size:12pt;font-weight:bold" align="center" | |- style="background-color:#8DB4E2;font-size:12pt;font-weight:bold" align="center" | ||
| width=" | | width="200" height="32" | Reagent | ||
| width=" | | width="120" | Single Rxn Vol | ||
| width=" | | width="120" | 14.2X MM uL | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
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|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| height="15" valign="bottom" | Bglii Cutter Primer (10 uM) | | height="15" valign="bottom" | Bglii Cutter Primer (10 uM) | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 5 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 71 | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | 10X Buffer 3.1 | | height="15" valign="bottom" | 10X Buffer 3.1 | ||
| align="center" align="center" valign="bottom" | 2 | | align="center" align="center" valign="bottom" | 2 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 28.4 | ||
|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| height="15" valign="bottom" | Bglii | | height="15" valign="bottom" | Bglii | ||
| align="center" align="center" valign="bottom" | 1 | | align="center" align="center" valign="bottom" | 1 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 14.2 | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | nfH2O | | height="15" valign="bottom" | nfH2O | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 7 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 99.4 | ||
|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| height="15" valign="bottom" | Total | | height="15" valign="bottom" | Total | ||
|style="font-weight:bold" align="center" align="center" valign="bottom" | 20 | |style="font-weight:bold" align="center" align="center" valign="bottom" | 20 | ||
|style="font-weight:bold" align="center" align="center" valign="bottom" | | |style="font-weight:bold" align="center" align="center" valign="bottom" | 213 | ||
|} | |} |
Revision as of 16:07, 21 March 2017
Circularization Comparison (T4 Poly/Ligase; Started Wednesday)
Bgl Digestion
Because I CAN. Also, will determine if there is an improvement in signal post-digestion.
Sample Matrix
Sample Origin | Condition | Sample Numbers |
F | T4 Ligase | 1,4 |
G | Phusion/Amp (Unpurified) | 1,4 |
H | Phusion/AmpHS (Unpurified) | 1,4 |
- Bglii Digestion
- Use samples 1 (Experiment) and 4 (No phosphate) for Bglii digest
- Make the following 4.2X master mix
- Aliquot 14 uL master mix into new tubes
- Add 5 uL appropriate sample to each tube
- Use the following thermocycler program
- 5 min 95C
- 10 min 50C
- 1 hr 37C; When this cycle starts add 1 uL Bglii
- Heat kill with 65C for 20 min
Reagent | Single Rxn Vol | 14.2X MM uL |
RCA Reaction Product | 5 | 0 |
Bglii Cutter Primer (10 uM) | 5 | 71 |
10X Buffer 3.1 | 2 | 28.4 |
Bglii | 1 | 14.2 |
nfH2O | 7 | 99.4 |
Total | 20 | 213 |