CTR:Notebook/Migrant/2017/03/28: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==Whole Genome Sequencing PCR-Free Library Prep for BUOW & COYE== | ||
* | * Used TruSeq Library Prep Kit | ||
*Diluted samples 0874-05557 (BUOW) & 98N2049 (COYE) to 1ug of DNA in 100 uL total of Low TE | |||
* Sonication (2017-04-10) | |||
**BioRuptor NGS: 4 cycles of 30 seconds on, 90 seconds off, High Power | |||
[[Image:BUOW COYE Sheared 04102017.JPG|100px]] | |||
1) 2uL sheared BUOW | |||
2) 2uL 100bp low scale ladder, | |||
3) 2uL sheared COYE | |||
→Sheared DNA to ~400bp. | |||
<br> | |||
* Dried down samples to get concentrated volume of 50 uL using SpeedVac (2017-04-10) | |||
* Clean-up fragmented DNA (2017-04-11) | |||
* End Repair and Size Selection- edited to size select for 400bp (2017-04-11) | |||
**Made a diluted bead mixture with the ratio of 109.25ul beads: 74.75ul dH<sub>2</sub>O and added 160 ul of mixture to each tube | |||
[[Image:BUOW COYE sizeselect 04112017.JPG|100px]] | |||
1) 2uL BUOW | |||
2) 2uL 100bp low scale ladder, | |||
3) 2uL COYE | |||
* Adenylate 3' Ends (2017-04-12) | |||
** Ran on JOHN thermocycler under program: SDCRKAD, Block B | |||
*Ligate Adapters (2017-04-12) | |||
**'''Assigned Adapter 3 to BUOW and Adapter 8 to COYE''' | |||
*Gel of Final Library: | |||
[[Image:BUOW COYE final 04122017.JPG|100px]] | |||
1) 2uL BUOW | |||
2) 2uL 100bp low scale ladder, | |||
3) 2uL COYE | |||
* Qubit: | |||
**'''BUOW: 7.72 ng/μL''' | |||
**'''COYE: 7.12 ng/μL''' | |||
*Run 1μL of sample + 2μL of dH<sub>2</sub>O on BioAnalyzer DNA High Sensitivity Chip (2017-04-12) | |||
**BUOW | |||
[[Image:BUOW WG Bioanalyzer.JPG|500px]] | |||
*COYE | |||
[[Image:COYE WG Bioanalyzer.JPG|500px]] | |||
*[http://www.openwetware.org/wiki/Image:BUOW_COYE_WG_Bioanalyzer_04122017.pdf Bioanalyzer Results] | |||
*Sent 10nM to Berkeley for sequencing (Illumina HiSeq 250 PER (Rapid Mode)) (2017-04-16) | |||
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__NOTOC__ | __NOTOC__ |
Latest revision as of 02:17, 27 September 2017
Project name | Main project page Previous entry Next entry |
Whole Genome Sequencing PCR-Free Library Prep for BUOW & COYE
1) 2uL sheared BUOW 2) 2uL 100bp low scale ladder, 3) 2uL sheared COYE
→Sheared DNA to ~400bp.
1) 2uL BUOW 2) 2uL 100bp low scale ladder, 3) 2uL COYE
1) 2uL BUOW 2) 2uL 100bp low scale ladder, 3) 2uL COYE
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