Daniel:Notebook/ComboLock/2017-3-13

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Revision as of 15:57, 13 March 2017 by >Djacobse (→‎Protocol)
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Bolt Test (Started Wednesday March 8)

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Protocol

Sample Matrix

Sample Condition
Sample 5 (AB) No Ligase
Sample 6 (AB) Normal
  1. Template-Bead Binding
    1. Suspend 2 uL (8 ug) beads per sample in 100 uL wash buffer
    2. Apply magnet for 30 sec and remove supernatant
    3. Add 2 uL 10 uM template oligo per sample to bead solution; incubate at RT for 5 min
    4. Wash beads with 100 uL wash buffer; vortex to suspend; apply magnet and remove supernatant
  2. C Probe Hybridization
    1. Combine following into a 0.2 mL tube
      1. 4 uL each PCCA-03 and PCCB-03 (10 uM each)
      2. 12 uL wash buffer
    2. Heat C probes to 90C for 5 min; Chill probes on ice to quench
    3. Add 5 uL probe mixture to beads
    4. Incubate at 40C for 1 hour with agitation
    5. Wash beads with 100 uL wash buffer; apply magnet and remove supernatant
    6. Repeat wash step above
    7. Repeat wash step above