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)

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Protocol-Part 2-qPCR

  1. qPCR
    1. Make qPCR master mixes according to following recipes
    2. 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
    3. Add 48 uL master mix to each well
    4. Add 2 uL sample according to table in following section
    5. qPCR Cycles
      1. 95C 3 min
      2. 95C 3 sec
      3. 55C 30 sec
      4. 72C 15 sec
      5. plate read
      6. goto b x24
      7. 72C 2 min
      8. 16C hold

    Plate Layouts

    File:20170209-Plate1-5.png

    File:20170209-Plate6-10.png