Team:ETH Zurich/labblog/20140702meet
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- | <html><article class=" | + | <html><article class="mini" id="20140702"></html> |
- | == | + | == Miniprep== |
- | = | + | = Wednesday, July 2nd = |
- | <html><section class=" | + | <html><section class="mini"></html> |
- | === | + | ===Miniprep piG0001 piG0029=== |
- | + | Addition of 200 μL Resuspension solution to culture pellet, vortex, addition of 200 μL Lysis solution <br/> | |
- | < | + | After ca. 5 min addition of 350 μL Neutralization solution, centrifuge samples for >10 min at 4000 rpm<br/> |
+ | Preparation of columns: addition of 500 μL Preparation solution, centrifugation for 1 min at 12 000 rcf, discard flow-through<br/> | ||
+ | Give supernatant of centrifuged samples onto columns, spin for 1 min at 12 000 rcf, discard flow-through<br/> | ||
+ | Addition of 500 μL Optional Wash Solution, spin for 1 min at 12 000 rcf, discard flow-through<br/> | ||
+ | Addition of 750 μL Wash Solution, spin for 1 min at 12 000 rcf, discard flow-through<br/> | ||
+ | Dry columns by centrifuging them for 2 min at 12 000 rcf, subsequently place columns in new collection tubes<br/> | ||
+ | Elute DNA with 100 μL Elution solution<br/> | ||
+ | According to Sigma-Aldrich Plasmid Miniprep Kit protocol<br/> | ||
- | + | Send samples piG0001, piG0015 and piG0029 for sequencing, primers used: oiG0001 for piG0001 and piG0020, oiG0002 for piG0015 | |
- | + | ||
- | + | ||
<html></section></html> | <html></section></html> | ||
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<html></article></html> | <html></article></html> |
Revision as of 09:48, 10 July 2014
Miniprep
Wednesday, July 2nd
Miniprep piG0001 piG0029
Addition of 200 μL Resuspension solution to culture pellet, vortex, addition of 200 μL Lysis solution
After ca. 5 min addition of 350 μL Neutralization solution, centrifuge samples for >10 min at 4000 rpm
Preparation of columns: addition of 500 μL Preparation solution, centrifugation for 1 min at 12 000 rcf, discard flow-through
Give supernatant of centrifuged samples onto columns, spin for 1 min at 12 000 rcf, discard flow-through
Addition of 500 μL Optional Wash Solution, spin for 1 min at 12 000 rcf, discard flow-through
Addition of 750 μL Wash Solution, spin for 1 min at 12 000 rcf, discard flow-through
Dry columns by centrifuging them for 2 min at 12 000 rcf, subsequently place columns in new collection tubes
Elute DNA with 100 μL Elution solution
According to Sigma-Aldrich Plasmid Miniprep Kit protocol
Send samples piG0001, piG0015 and piG0029 for sequencing, primers used: oiG0001 for piG0001 and piG0020, oiG0002 for piG0015