Matt:LabNotes/2014-9-6

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Revision as of 21:55, 6 September 2014 by >Mzcai (Created page with "=="Tertiary" Rolony Synthesis== *Hypothesis: We can increase the number of gene-specific rolonies (previously known as secondary rolonies) by repeating padlock probe capture a...")
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"Tertiary" Rolony Synthesis

  • Hypothesis: We can increase the number of gene-specific rolonies (previously known as secondary rolonies) by repeating padlock probe capture and rolonization on samples with primary and secondary rolonies
    • There should be more targets for the padlock probes to capture since all primary rolonies are still fixed and 100% of secondary rolonies will be possible targets for padlock probes
    • May increase amplification bias

Procedure

  • Add 100ul Padlock Probe Mix
Components Volume
5 U/ul Ampligase 10
10X Ampligase Buffer 10
908 nM Agi26k0gap 18.6
H2O 61.4
Total 100
  • Incubate at 37C for 30min
  • Incubate at 45C for 24hrs