CTR:Notebook/PIRE/Entry Base: Difference between revisions
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* Clean up (2014-04-30) | * Clean up (2014-04-30) | ||
**Take 5 uL | **Take 5 uL from each well and combine to 1.7 mL tube | ||
**AMPure bead clean up: | **AMPure bead clean up: | ||
***Use 480 uL beads to DNA (1:1) | ***Use 480 uL beads to DNA (1:1) | ||
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****72 degrees for 30 seconds | ****72 degrees for 30 seconds | ||
***72 degrees for 5 minutes | ***72 degrees for 5 minutes | ||
[[Image:Sunbirds1 PCR 2014-05-01.JPG|200px]] | |||
1) 1 uL DNA template, 2) 2 uL 100 bp low scale ladder, 3) 5 uL PCR product | |||
*Bead clean up (2014-05-02) | |||
**Use 45 uL AMPure beads (1:1) | |||
**2 washes of 200 uL 80% EtOH | |||
**Elute in 30 uL LowTE | |||
*Quant DNA library using PicoGreen and run (<3 ng/uL) on BioAnalyzer DNA High Sensitivity Chip | |||
[[Image:Sunbirds1 bioanalyzer.jpg|450px]] | |||
*Dilute library to 10 nM in 10 uL and send to Berkeley for sequencing (Illumina HiSeq 100 SR) (2014-05-05) | |||
Revision as of 14:07, 21 November 2014
PIRE RAD Library Preps | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page |
RAD Library Prep - Olive Sunbirds Plate 1
1) 1uL pre-sheared DNA, 2) 2uL 100bp low scale ladder, 3) 3uL sheared DNA
1) 1 uL DNA template, 2) 2 uL 100 bp low scale ladder, 3) 5 uL PCR product
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