Andrew:Notebook/C1 OpenApp 150421: Difference between revisions

From ZhangLabWiki
Jump to navigation Jump to search
>Andrew
(Created page with "400px")
 
>Andrew
No edit summary
 
(17 intermediate revisions by the same user not shown)
Line 1: Line 1:
[[File:C1_OpenApp_150421.jpg|400px]]
==C1_OpenApp_150421==
 
===Notes===
*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===
 
====Overview====
 
[[File:Reagents-page-002.jpg|800px]]
 
====Reagents====
 
[[File:Reagents-page-001.jpg|800px]]
 
[[File:Reagents.xlsx|Reagent excel file]]
 
====Protocol====
 
[[File:Protocol_Worksheet_HLS_150421_v1-page-001.jpg|800px]]
 
[[File:Protocol_Worksheet_HLS_150421_v1-page-002.jpg|800px]]
 
[[File:Protocol_Worksheet_HLS_150421_v1-page-003.jpg|800px]]
 
===Screening===
 
====Size Select====
[[File:C1_OpenApp_150421.jpg|800px]]
 
===Re-sequencing===
 
====Size Select====
 
[[File:C1_OpenApp_150421_gel2.jpg|800px]]

Latest revision as of 23:57, 6 May 2015

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