Daniel:Notebook/ComboLock/2017-2-10: Difference between revisions
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<li>Make qPCR master mixes according to following recipes</li> | <li>Make qPCR master mixes according to following recipes</li> | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#B1A0C7;font-size:12pt;font-weight:bold" align="center" | |||
| width="1005" height="45" | Master Mix | |||
| width="85" | No X | |||
| width="190" | Primer Pair | |||
| width="82" | Forward Primer Amt (10 uM) | |||
| width="80" | Reverse Primer Amt (10 uM) | |||
| width="65" | uL nfH2O | |||
| width="65" | uL 2X Kapa SYBR Fast | |||
| width="65" | Total | |||
|- style="font-size:12pt" | |||
| height="15" valign="bottom" | MMC1 | |||
| align="center" valign="bottom" | 73 | |||
| align="center" valign="bottom" | Primer24/Primer4RC | |||
| align="center" align="center" valign="bottom" | 73 | |||
| align="center" align="center" valign="bottom" | 73 | |||
| align="center" align="center" valign="bottom" | 1168 | |||
| align="center" align="center" valign="bottom" | 1460 | |||
|style="font-weight:bold" align="center" align="center" valign="bottom" | 2774 | |||
|- style="background-color:#BFBFBF;font-size:12pt" | |||
| height="15" valign="bottom" | MMC2 | |||
| align="center" valign="bottom" | 2.2 | |||
| align="center" valign="bottom" | Primer6/Primer12RC | |||
| align="center" align="center" valign="bottom" | 2.2 | |||
| align="center" align="center" valign="bottom" | 2.2 | |||
| align="center" align="center" valign="bottom" | 35.2 | |||
| align="center" align="center" valign="bottom" | 44 | |||
|style="font-weight:bold" align="center" align="center" valign="bottom" | 83.6 | |||
|- style="font-size:12pt" | |||
| height="15" valign="bottom" | MMT | |||
| align="center" valign="bottom" | 2.2 | |||
| align="center" valign="bottom" | PCCTAmpF/PCCT-AmpR | |||
| align="center" align="center" valign="bottom" | 2.2 | |||
| align="center" align="center" valign="bottom" | 2.2 | |||
| align="center" align="center" valign="bottom" | 35.2 | |||
| align="center" align="center" valign="bottom" | 44 | |||
|style="font-weight:bold" align="center" align="center" valign="bottom" | 83.6 | |||
|} | |||
<li>Add 48 uL master mix to each well</li> | <li>Add 48 uL master mix to each well</li> |
Revision as of 01:14, 10 February 2017
Beads Binding Test (Started Yesterday)
Protocol-Part 2-qPCR
- qPCR
- Make qPCR master mixes according to following recipes
- Add 48 uL master mix to each well
- Add 2 uL sample according to table in following section
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 15 sec
- plate read
- goto b x24
- 72C 2 min
- 16C hold
Master Mix | No X | Primer Pair | Forward Primer Amt (10 uM) | Reverse Primer Amt (10 uM) | uL nfH2O | uL 2X Kapa SYBR Fast | Total |
MMC1 | 73 | Primer24/Primer4RC | 73 | 73 | 1168 | 1460 | 2774 |
MMC2 | 2.2 | Primer6/Primer12RC | 2.2 | 2.2 | 35.2 | 44 | 83.6 |
MMT | 2.2 | PCCTAmpF/PCCT-AmpR | 2.2 | 2.2 | 35.2 | 44 | 83.6 |