Matt:LabNotes/2016-3-31: Difference between revisions

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(Created page with "=Bead with PKP2 Target for DARTFISH Positive Control== *Using PA gel may prevent diffusion of probes and enzymes during DARTFISH *To test this and to have a positive control f...")
 
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==Capture with CA12kNov2014 V4 in tube==
==Capture with CA12kNov2014 V4 in tube==
[[Matt:LabNotes/2015-5-18 | Reference protocol]]


==PCR + Sequencing Adapters==
==PCR + Sequencing Adapters==

Revision as of 01:35, 8 April 2016

Bead with PKP2 Target for DARTFISH Positive Control=

  • Using PA gel may prevent diffusion of probes and enzymes during DARTFISH
  • To test this and to have a positive control for future experiments need to design a synthetic target
    • Attach target to magnetic streptavidin bead
  • Design oligonucleotide with biotin 5' modification
    • Chose PKP2 gene because haven't detected it in any DARTFISH samples
    • Also this specific padlock probe has very high efficiency when measured in tube
 PKP2_control	/5BiosG/AAAAAAGAGATGGCTGTCTTTTTCACACTTGGGTCACCAACATGCAGCATCTTTC

Link PKP2_control oligo to Streptavidin Bead

Measure DNA Concentration

Capture with CA12kNov2014 V4 in tube

Reference protocol

PCR + Sequencing Adapters