Daniel:Notebook/ComboLock/2017-3-30: Difference between revisions

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=RCA Test (Started [[Daniel:Notebook/ComboLock/2017-3-24|March 24]])=
=RCA Test (Started [[Daniel:Notebook/ComboLock/2017-3-24|March 24]])=
<ol>
<li>Qiaquick Column</li>
<ol type="A">
<li>Add 100 uL PB (binding buffer) to sample</li>
<li>Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through</li>
<li>Add 700 uL PE (wash buffer) to column and spin for 1 minute at 14000 rpm; discard flow through</li>
<li>Dry spin column for 2 minutes at 14000 rpm; discard flow through</li>
<li>Let stand with cover open in fume hood for ~5 minutes</li>
<li>Transfer column to a new 1.5 mL eppendorf tube</li>
<li>Add ≥30 uL EB to column</li>
<li>Let stand 1 minute</li>
<li>Spin for 1 minute at 14000 rpm</li>
</ol>
<li>TBE Gel</li>
<ol type="A">
<li>Mix 72 uL TBE and 24 uL 6X dye</li>
<li>Aliquot 8 uL on to parafilm per sample/ladder</li>
<li>Add 4 uL reaction or 2 uL ladder to appropriate aliquot</li>
<li>Add 10 uL mix to gel lanes</li>
<li>Run gel for 24 minutes at 230V</li>
<li>Stain with 2 uL SYBR gold for 3 minutes</li>
<li>Rinse and image in gel doc</li>
</ol>
</ol>


[[Daniel:Notebook/ComboLock|Back to Calendar]]
[[Daniel:Notebook/ComboLock|Back to Calendar]]

Revision as of 22:48, 30 March 2017

RCA Test (Started March 24)

  1. Qiaquick Column
    1. Add 100 uL PB (binding buffer) to sample
    2. Load sample onto column and spin for 1 minute at 14000 rpm; discard flow through
    3. Add 700 uL PE (wash buffer) to column and spin for 1 minute at 14000 rpm; discard flow through
    4. Dry spin column for 2 minutes at 14000 rpm; discard flow through
    5. Let stand with cover open in fume hood for ~5 minutes
    6. Transfer column to a new 1.5 mL eppendorf tube
    7. Add ≥30 uL EB to column
    8. Let stand 1 minute
    9. Spin for 1 minute at 14000 rpm
  2. TBE Gel
    1. Mix 72 uL TBE and 24 uL 6X dye
    2. Aliquot 8 uL on to parafilm per sample/ladder
    3. Add 4 uL reaction or 2 uL ladder to appropriate aliquot
    4. Add 10 uL mix to gel lanes
    5. Run gel for 24 minutes at 230V
    6. Stain with 2 uL SYBR gold for 3 minutes
    7. Rinse and image in gel doc

Back to Calendar