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> | ||
'''Master Mix Recipes''' | |||
{| 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:#B1A0C7;font-size:12pt;font-weight:bold" align="center" | |- style="background-color:#B1A0C7;font-size:12pt;font-weight:bold" align="center" | ||
| width=" | | width="65" height="45" | Master Mix NoX | ||
| width=" | | width="65" | Primer Pair | ||
| width="65" | Forward Primer Amt (10 uM) | |||
| width=" | | width="65" | Reverse Primer Amt (10 uM) | ||
| width=" | |||
| width="65" | uL nfH2O | | width="65" | uL nfH2O | ||
| width=" | | width="65" | uL 2X Kapa SYBR Fast | ||
| width="65" | Total | | width="65" | Total | ||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="30" | 73 | |||
| Primer24/Primer4RC | |||
| align="center" | 73 | |||
| align="center" | 73 | |||
| align="center" | 1168 | |||
| align="center" | 1460 | |||
|style="font-weight:bold" align="center" | 2774 | |||
|- style="background-color:#BFBFBF;font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="30" | 63 | |||
| Primer6/Primer12RC | |||
| align="center" | 63 | |||
| align="center" | 63 | |||
| align="center" | 1008 | |||
| align="center" | 1260 | |||
|style="font-weight:bold" align="center" | 2394 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="30" | 2.2 | |||
| PCCTAmpF/PCCT-AmpR | |||
| align="center" | 2.2 | |||
| align="center" | 2.2 | |||
| align="center" | 35.2 | |||
| align="center" | 44 | |||
|style="font-weight:bold" align="center" | 83.6 | |||
|} | |||
'''Plate Organization''' | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#DA9694;font-size:12pt" align="center" | |||
| width="65" height="28" | Plate | |||
| width="65" | Master Mix Quantity | |||
| width="65" | Samples | |||
| width="65" | Target | |||
| width="138" | Primer Pair | |||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | | | height="15" valign="bottom" | 1 | ||
| align="center" align="center" valign="bottom" | 73 | | align="center" align="center" valign="bottom" | 73 | ||
| align="center" valign="bottom" | 1-5 | |||
| align="center" valign="bottom" | Template | |||
| align="center" valign="bottom" | CLv3-AmpF/CLv3-AmpR | |||
|- style="background-color:#BFBFBF;font-size:12pt" | |||
| height="15" valign="bottom" | 2 | |||
| align="center" align="center" valign="bottom" | 63 | |||
| align="center" valign="bottom" | 6-10 | |||
| align="center" valign="bottom" | Template | |||
| align="center" valign="bottom" | CLv3-AmpF/CLv3-AmpR | |||
|- style="font-size:12pt" | |||
| height="15" valign="bottom" | 3 | |||
| align="center" align="center" valign="bottom" | 73 | | align="center" align="center" valign="bottom" | 73 | ||
| | | align="center" valign="bottom" | 1-5 | ||
| | | align="center" valign="bottom" | C1 | ||
| | | align="center" valign="bottom" | primer24/primer4RC | ||
|- style="background-color:#BFBFBF;font-size:12pt" | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| height="15" valign="bottom" | | | height="15" valign="bottom" | 4 | ||
| align="center" | | align="center" align="center" valign="bottom" | 63 | ||
| align="center" valign="bottom" | 6-10 | |||
| | | align="center" valign="bottom" | C1 | ||
| | | align="center" valign="bottom" | primer24/primer4RC | ||
| | |||
|- style="font-size:12pt" | |- style="font-size:12pt" | ||
| height="15" valign="bottom" | | | height="15" valign="bottom" | 5 | ||
| align="center" | | align="center" align="center" valign="bottom" | 73 | ||
| align="center" valign="bottom" | | | align="center" valign="bottom" | 1-5 | ||
| align="center" align="center" valign="bottom" | | | align="center" valign="bottom" | C2 | ||
| | | align="center" valign="bottom" | primer6/primer12RC | ||
| align="center" align="center" valign="bottom" | | |||
| | |- style="background-color:#BFBFBF;font-size:12pt" | ||
| | | height="15" valign="bottom" | 6 | ||
| align="center" align="center" valign="bottom" | 63 | |||
| align="center" valign="bottom" | 6-10 | |||
| align="center" valign="bottom" | C2 | |||
| align="center" valign="bottom" | primer6/primer12RC | |||
|} | |} |
Revision as of 19:28, 10 February 2017
Beads Binding Test (Started Yesterday)
Protocol-Part 2-qPCR
- qPCR
- Make qPCR master mixes according to following recipes Master Mix 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 NoX | Primer Pair | Forward Primer Amt (10 uM) | Reverse Primer Amt (10 uM) | uL nfH2O | uL 2X Kapa SYBR Fast | Total |
73 | Primer24/Primer4RC | 73 | 73 | 1168 | 1460 | 2774 |
63 | Primer6/Primer12RC | 63 | 63 | 1008 | 1260 | 2394 |
2.2 | PCCTAmpF/PCCT-AmpR | 2.2 | 2.2 | 35.2 | 44 | 83.6 |
Plate Organization
Plate | Master Mix Quantity | Samples | Target | Primer Pair |
1 | 73 | 1-5 | Template | CLv3-AmpF/CLv3-AmpR |
2 | 63 | 6-10 | Template | CLv3-AmpF/CLv3-AmpR |
3 | 73 | 1-5 | C1 | primer24/primer4RC |
4 | 63 | 6-10 | C1 | primer24/primer4RC |
5 | 73 | 1-5 | C2 | primer6/primer12RC |
6 | 63 | 6-10 | C2 | primer6/primer12RC |
Plate Layouts
A Plate (Samples 1-5)
B Plate (Samples 6-10)