Daniel:Notebook/ComboLock/2016-9-26: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Djacobse
(Created page with "=PCR Testing (Started Thursday)= Back to Calendar Continuing the tests on qPCR for the positive control...")
 
>Djacobse
Line 14: Line 14:
{| 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:#B8CCE4;font-size:12pt;font-weight:bold" align="center"
|- style="background-color:#B8CCE4;font-size:12pt;font-weight:bold" align="center"
| width="65" height="45" | Sample
| width="105" height="45" | Sample
| width="65" | Molecule Count
| width="65" | Molecule Count
| width="65" | Lane
| width="65" | Lane

Revision as of 18:30, 26 September 2016

PCR Testing (Started Thursday)

Back to Calendar

Continuing the tests on qPCR for the positive control amplicon using primer concentrations. This is following some of the conclusions from Friday.

Protocol

  1. qPCR
    1. Follow reaction map below
    2. Sample Molecule Count Lane AmpR Index Sample Vol (uL) 2X Kapa SYBR qPCR MM 10 uM AmpF 6.4 10 uM AmpR6.3-IndXX H2O Total Volume (uL) Master Mix
      Sample 1A 1.00E+10 A1 36 2 25 5 5 13 50 MM1
      Sample 1B 1.00E+10 A2 36 2 25 5 5 13 50 MM1
      Sample 1C 1.00E+10 A3 36 2 25 5 5 13 50 MM1
      NTC 0 A4 36 0 25 5 5 13 50 NTC
      Sample 1A 1.00E+09 A5 36 2 25 1 1 21 50 MM2
      Sample 1B 1.00E+09 A6 36 2 25 1 1 21 50 MM2
      Sample 1C 1.00E+09 A7 36 2 25 1 1 21 50 MM2
      NTC 0 A8 36 0 25 1 1 19.5 50 NTC
      Sample 1A 1.00E+10 H1 36 2 25 0.5 0.5 22 50 MM3
      Sample 1B 1.00E+10 H2 36 2 25 0.5 0.5 22 50 MM3
      Sample 1C 1.00E+10 H3 36 2 25 0.5 0.5 22 50 MM3
      NTC 0 H4 36 0 25 0.5 0.5 19.5 50 NTC
      Sample 1A 1.00E+09 H5 37 2 25 0.1 0.1 22.8 50 MM4
      Sample 1B 1.00E+09 H6 37 2 25 0.1 0.1 22.8 50 MM4
      Sample 1C 1.00E+09 H7 37 2 25 0.1 0.1 22.8 50 MM4
      NTC 0 H8 37 0 25 0.1 0.1 19.5 50 NTC
      Sample 1A 1.00E+10 C1 37 2 25 0.01 0.01 23 50 MM5
      Sample 1B 1.00E+10 C2 37 2 25 0.01 0.01 23 50 MM5
      Sample 1C 1.00E+10 C3 37 2 25 0.01 0.01 23 50 MM5
      NTC 0 C4 37 0 25 0.01 0.01 19.5 50 NTC
    3. Add 48 uL correct master mix to each well
    4. qPCR Cycles
      1. 95C 3 min
      2. 95C 3 sec
      3. 55C 30 sec
      4. 72C 20 sec
      5. plate read
      6. goto b x26
      7. 72C 2 min
      8. 16C hold