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

From ZhangLabWiki
Jump to navigation Jump to search
>Djacobse
>Djacobse
Line 28: Line 28:
<li>Use "standard" thermocycling program</li>
<li>Use "standard" thermocycling program</li>
</ol>
</ol>
==Results==
<gallery perrow=2 heights=250px widths=350px mode=packed-hover caption="Single Biotin Results">
File:PlateLayout-20170613-BiotinTest.png|Plate array
File:Singlebiotin-20170613-plateCTs.png|Plate CTs
File:Singlebiotin-20170613-plateCTs-normed.png|Normalized plate CTs
File:Singlebiotin-20170613-rawcurves.png|Raw curves
</gallery>
<gallery perrow=2 heights=250px widths=350px mode=packed-hover caption="Single Biotin Results">
File:PlateLayout-20170613-BiotinTest.png|Plate array
File:Dualbiotin-20170613-plateCTs.png|Plate CTs
File:Dualbiotin-20170613-plateCTs-normed.png|Normalized plate CTs
File:Dualbiotin-20170613-rawcurves.png|Raw curves
</gallery>

Revision as of 17:01, 14 June 2017

Biotin vs Dual Biotin (Started Yesterday)

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 layout (4X master mix is for positive controls, 58X is for samples and negative controls)
    4. File:PlateLayout-20170613-BiotinTest.png
    5. Add 2 uL appropriate sample according to plate layout
    6. Use "standard" thermocycling program

    Results