Andrew:Notebook/C1 OpenApp 150421

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C1_OpenApp_150421[edit]

Notes[edit]

  • This protocol uses HLS/Protease followed by Tn5-059
  • 25 X Tn5-059 is 17.5 uM --> 1 X = 700 nM
  • Illumina has been using 40 nM = .057 X Tn5-059 on 5 ng DNA --> 7e-5 X for 6 pg
  • I will use 0.1 X Tn5-059 based on previous experiments
  • PCR with Illumina NPM from Nextera kit
  • PCR mix loaded into chamber 4, with barcodes backloaded into chamber 5
  • Sample: AD 18-99 Full PI NeuN+ cortex sorted 4/8/2015 frozen at 350/uL in 5 uL aliquots

Setup[edit]

Overview[edit]

File:Reagents-page-002.jpg

Reagents[edit]

File:Reagents-page-001.jpg

Reagent excel file

Protocol[edit]

File:Protocol Worksheet HLS 150421 v1-page-001.jpg

File:Protocol Worksheet HLS 150421 v1-page-002.jpg

File:Protocol Worksheet HLS 150421 v1-page-003.jpg

Screening[edit]

Size Select[edit]

File:C1 OpenApp 150421.jpg

Re-sequencing[edit]

Size Select[edit]

File:C1 OpenApp 150421 gel2.jpg