Daniel:Notebook/PosSequencing/2017-6-13: Difference between revisions

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<li>Prepare 2 plates; 1 single biotin and 1 dual biotin</li>
<li>Prepare 2 plates; 1 single biotin and 1 dual biotin</li>
<li>For each sample prepare 58X master mix and (4X) master mix: 1X components set to 30 uL; add 2uL sample</li>
<li>For each sample prepare 58X master mix and (4X) master mix: 1X components set to 30 uL; add 2uL sample</li>
<li>ol type="a">
<ol type="a">
<li>58 (4) uL 1 uM primer2</li>
<li>58 (4) uL 1 uM primer2</li>
<li>(4) uL 1 uM biotin/2biotin uracil primer</li>
<li>(4) uL 1 uM biotin/2biotin uracil primer</li>

Revision as of 00:02, 13 June 2017

Biotin vs Dual Biotin

Back to Calendar


Workflow Reminder

File:ExperimentMap-20170610-Biotin-DualBiotin.png

Protocol

  1. qPCR
    1. Prepare 2 plates; 1 single biotin and 1 dual biotin
    2. For each sample prepare 58X master mix and (4X) master mix: 1X components set to 30 uL; add 2uL sample
      1. 58 (4) uL 1 uM primer2
      2. (4) uL 1 uM biotin/2biotin uracil primer
      3. 58 uL 10 pM barcode oligo v2
      4. 870 (60) uL 2X Kapa SYBR
      5. 638 (44) uL nfH2O
    3. Add 28 uL master mix according to plate (4X master mix is for positive controls, 58X is for samples and negative controls)
    4. Add 2 uL appropriate sample according to plate layout
    5. Use "standard" thermocycling program