Andrew:Notebook/C1 OpenApp 150421: Difference between revisions
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[[File:C1_OpenApp_150421.jpg| | ==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]
Reagents[edit]
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