Daniel:Notebook/ComboLock/2017-3-9: Difference between revisions
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|style="background-color:#66CCFF;font-weight:bold" width="85" | MMC1 (11.2X) | |style="background-color:#66CCFF;font-weight:bold" width="85" | MMC1 (11.2X) | ||
|style="background-color:#66CCFF;font-weight:bold" width="85" | MMC2 (11.2X) | |style="background-color:#66CCFF;font-weight:bold" width="85" | MMC2 (11.2X) | ||
|style="background-color:#66CCFF;font-weight:bold" width="85" | MMP ( | |style="background-color:#66CCFF;font-weight:bold" width="85" | MMP (12.2X) | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
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| align="center" align="center" valign="bottom" | 11.2 | | align="center" align="center" valign="bottom" | 11.2 | ||
| align="center" align="center" valign="bottom" | 11.2 | | align="center" align="center" valign="bottom" | 11.2 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 12.2 | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
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| align="center" align="center" valign="bottom" | 11.2 | | align="center" align="center" valign="bottom" | 11.2 | ||
| align="center" align="center" valign="bottom" | 11.2 | | align="center" align="center" valign="bottom" | 11.2 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 12.2 | ||
|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
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| align="center" align="center" valign="bottom" | 235.2 | | align="center" align="center" valign="bottom" | 235.2 | ||
| align="center" align="center" valign="bottom" | 235.2 | | align="center" align="center" valign="bottom" | 235.2 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 256.2 | ||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
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| align="center" align="center" valign="bottom" | 280 | | align="center" align="center" valign="bottom" | 280 | ||
| align="center" align="center" valign="bottom" | 280 | | align="center" align="center" valign="bottom" | 280 | ||
| align="center" align="center" valign="bottom" | | | align="center" align="center" valign="bottom" | 305 | ||
|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
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|style="font-weight:bold" align="center" align="center" valign="bottom" | 537.6 | |style="font-weight:bold" align="center" align="center" valign="bottom" | 537.6 | ||
|style="font-weight:bold" align="center" align="center" valign="bottom" | 537.6 | |style="font-weight:bold" align="center" align="center" valign="bottom" | 537.6 | ||
|style="font-weight:bold" align="center" align="center" valign="bottom" | | |style="font-weight:bold" align="center" align="center" valign="bottom" | 585.6 | ||
|} | |} |
Revision as of 19:09, 9 March 2017
Bolt Test (Started Yesterday)
Protocol
- Bolt/Latch Hybridization
- Wash twice with 100 uL wash buffer
- Elution
- Add 50 uL wash buffer to each sample
- Cook at 95C for 15 minutes
- qPCR
- Make master mixes according to following recipes
- Add 48 uL master mix to each well
- Add 2 uL sample according to plate layout File:PlateLayout-20170309-BoltTest.png
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x30
- 72C 2 min
- 16C hold
- TBE Gel
- Mix 96 uL TBE, 24 uL 6x loading dye
- Aliquot 10 uL per sample/ladder lane onto parafilm
- Add 2 uL of sample or ladder to correct drop
- Load 10 uL in to well
- Run gel for 23 minutes at 230V
- Open gel and stain with 3 uL SYBR Gold for 3 minutes
- Rinse gel and image in gel doc
Single Rxn | MMC1 (11.2X) | MMC2 (11.2X) | MMP (12.2X) | |
Primer Pair | NA | p24/p4RC | p6/p12RC | p24/p12RC |
uL Fwd Primer (10 uM) | 1 | 11.2 | 11.2 | 12.2 |
uL Rev Primer (10 uM) | 1 | 11.2 | 11.2 | 12.2 |
uL nfH2O | 21 | 235.2 | 235.2 | 256.2 |
uL Kapa SYBR Fast | 25 | 280 | 280 | 305 |
Total | 48 | 537.6 | 537.6 | 585.6 |