Matt:LabNotes/2015-3-25: Difference between revisions

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>Mzcai
(Created page with "==CA12k_Nov2014_V4 Probe Production== *Production of Padlock Probes (V4) Matt:LabNotes/2014-11-19#Assembly_of_oligo_pool_for_ordering *Do double 96 100ul PCR Reactions ==...")
 
>Mzcai
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'''Program''' 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold
'''Program''' 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold
[[File: | 800px]]
[[File:032515_CA12kNov14_ProductionPCR_V4.JPG | 800px]]
 
===Zymo DNA Clean & Concentrator 100===
*Purify 10ml of PCR product with 1 Zymo-spin V Column/Reservoir
**Add 50ml DNA Binding Buffer to 10ml product (Use 2 50ml tubes)
*Elute with 540ul H2O
*Measured concentration with Nanodrop:
~540ul of V4 probes: __ ng/ul => ug
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===Qia Column Purification===
===Qia Column Purification===

Revision as of 20:38, 25 March 2015

CA12k_Nov2014_V4 Probe Production

Production PCR

Components Volume (1X) Volume (100X)
First round amplicon CA12k_Nov14_V7 (10nM) 0.2 20
2X KAPA SYBG MM 50 5000
AP1V4U (100uM) 0.4 40
AP2V4 (100uM) 0.4 40
H2O 49 4900
Total 100 10000

Program 98C 1min -> (98C 10sec -> 55C 30sec -> 72C 45sec) x 15 -> 72C 2min -> 15C hold File:032515 CA12kNov14 ProductionPCR V4.JPG

Zymo DNA Clean & Concentrator 100

  • Purify 10ml of PCR product with 1 Zymo-spin V Column/Reservoir
    • Add 50ml DNA Binding Buffer to 10ml product (Use 2 50ml tubes)
  • Elute with 540ul H2O
  • Measured concentration with Nanodrop:

~540ul of V4 probes: __ ng/ul => ug