CTR:Notebook/PIRE/2014/07/09: Difference between revisions
Sirena Lao (talk | contribs) |
(fix raw html notebook nav) |
||
(3 intermediate revisions by one other user not shown) | |||
Line 2: | Line 2: | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> PIRE RAD Library Preps</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> PIRE RAD Library Preps</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}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
Line 148: | Line 148: | ||
==Re-prep Greenbuls Plate 2== | ==Re-prep Greenbuls Plate 2== | ||
Used same SbfI-HF and ligase from previous library. | |||
*New DNA plate prepped 2014-08-05 | *New DNA plate prepped 2014-08-05 | ||
*Digestion: RADIGEST on Tony 2014-08-06 | *Digestion: RADIGEST on Tony 2014-08-06 | ||
Line 186: | Line 188: | ||
3) 2 μL 100bp ladder; | 3) 2 μL 100bp ladder; | ||
4) 1 μL Greenbul plate 2 template; | 4) 1 μL Greenbul plate 2 template; | ||
5) 5 μL Greenbul plate 2 | 5) 5 μL Greenbul plate 2 PCR product | ||
→Using new SbfI-HF seems to work. New enzyme and ligase is ideal! | →Using new SbfI-HF seems to work. New enzyme and ligase is ideal! | ||
Line 193: | Line 195: | ||
**Sunbirds Plate 2: 4.74 ng/μL | **Sunbirds Plate 2: 4.74 ng/μL | ||
**Greenbuls Plate 2: 7.79 ng/μL | **Greenbuls Plate 2: 7.79 ng/μL | ||
==BioAnalyzer results from all 3 libraries== | |||
1) Sunbirds Plate 2; 2) Greenbuls Plate 1; 3) Greenbuls Plate 2 | |||
[[Image:Bioanalyzer OS2LG12.png|450px]] | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Latest revision as of 00:09, 27 September 2017
PIRE RAD Library Preps | Main project page Previous entry Next entry |
RAD Library Prep - Olive Sunbirds Plate 2, Little Greenbuls Plates 1 & 2
2μL of sheared DNA in each well
Troubleshooting
1) Pre-cleaned DNA (post-P1-ligation), 2) Cleaned DNA, 3) Sheared DNA
1) 1μL template, 2) 5μL PCR product
1&2) Sunbirds Plate 1, 3&4) Greenbuls Plate 1 (from re-sheared template) Re-prep Greenbuls Plate 1 using leftover DNA from plateUsed new SbfI-HF tube and new T4 ligase. 2014-07-31:
2014-08-01:
1μL template, 2μL 100bp ladder, 5μL PCR product
Re-prep Greenbuls Plate 2Used same SbfI-HF and ligase from previous library.
2014-08-07:
8 cycles of 15 seconds on, 90 seconds off, high power. 2μL of ladder and sheared DNA.
1) 1μL template. 2) 2μL ladder. 3) 5μL PCR product Re-prep Sunbirds Plate 2 and Greenbuls Plate 2Re-prep from existing plates. Received new SbfI-HF from NEB.
1) 1 μL Sunbird plate 2 template; 2) 5 μL Sunbird plate 2 PCR product; 3) 2 μL 100bp ladder; 4) 1 μL Greenbul plate 2 template; 5) 5 μL Greenbul plate 2 PCR product →Using new SbfI-HF seems to work. New enzyme and ligase is ideal!
BioAnalyzer results from all 3 libraries1) Sunbirds Plate 2; 2) Greenbuls Plate 1; 3) Greenbuls Plate 2
|