CTR:Notebook/PIRE/2014/07/09: Difference between revisions
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→ PCR products do not look 2x as bright as the template. No significant amplication. | → PCR products do not look 2x as bright as the template. No significant amplication. | ||
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==Troubleshooting== | ==Troubleshooting== | ||
*Tried re-prep of leftover sheared DNA (Blunt End Repair → PCR), but no difference | *Tried re-prep of leftover sheared DNA (Blunt End Repair → PCR), but no difference | ||
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→ PCR with good template still works. P2 primer only does not work at all | → PCR with good template still works. P2 primer only does not work at all | ||
==Re-prep Greenbuls Plate 1 using leftover DNA from plate== | |||
2014-07-31: | |||
*Digestion: RADIGEST on Tony | |||
*Ligation: RADLIG on Tony | |||
**Used adapters from original Berkeley plate (undiluted) | |||
*Clean up | |||
*Sonication: 8 cycles of 15 seconds on, 90 seconds off, High Power | |||
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Revision as of 12:44, 31 July 2014
PIRE RAD Library Preps | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
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 plate2014-07-31:
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