User:Daniel Goodman/Notebook/Cluzel/2010/04/05: Difference between revisions
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* Vector Nomenclature: http://www.expressys.com/main_vectors.html | * Vector Nomenclature: http://www.expressys.com/main_vectors.html | ||
===Electroporations=== | |||
{| class="wikitable" border="1" | {| class="wikitable" border="1" | ||
|- | |- | ||
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| 1.62 | | 1.62 | ||
|} | |} | ||
Doing 4 electroporations, in order to ensure competent cells made last week work correctly. | Doing 4 electroporations, in order to ensure competent cells made last week work correctly. | ||
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{| class="wikitable" border="1" | {| class="wikitable" border="1" | ||
|- | |- | ||
! Symbol | |||
! Media | ! Media | ||
! Plasmid | ! Plasmid | ||
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! Why | ! Why | ||
|- | |- | ||
| A | |||
| LA/Amp | | LA/Amp | ||
| PZE-12-CFP | | PZE-12-CFP | ||
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| To get comp cells w/ plasmid | | To get comp cells w/ plasmid | ||
|- | |- | ||
| B | |||
| LA | | LA | ||
| PZE-12-CFP | | PZE-12-CFP | ||
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| To check that my competent cells are alive | | To check that my competent cells are alive | ||
|- | |- | ||
| | | C | ||
| | | Spec/AMP | ||
| PZS4-ATTP | |||
| My competent MG1655 | | My competent MG1655 | ||
| To check that my competent cells transform | | To check that my competent cells transform | ||
|- | |- | ||
| D | |||
| LA/AMP | | LA/AMP | ||
| PZE-12-CFP | | PZE-12-CFP | ||
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|} | |} | ||
Protocol steps on page 1.119 - 1.122 of Molecular Cloning, Volume 1 book | |||
* used straight glycerol instead of GYT | |||
* used SOC medium (1 ml per reaction) | |||
* 1 ul of DNA per experiment (approx 52 ng for my plasmid, did not nanodrop Jeff's spec plasmid) | |||
* 1.8 kV, 1 second shock | |||
Put in 37 degree shaker w/ the SOC for 1 hour. Spun down, supernatant removed to ~50ul each, resuspended, plated and spun onto correct antibiotic plates w/ LB. Put in 37 degree incubator overnight at 7:45 pm. Will take out tomorrow morning. | |||
|} | |} | ||
__NOTOC__ | __NOTOC__ |
Latest revision as of 16:48, 5 April 2010
Cluzel Lab Notebook Daniel Goodman |
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Prepping Plasmid for ElectroporationPZE-12-CFP vector from JM Kim
Electroporations
Doing 4 electroporations, in order to ensure competent cells made last week work correctly.
Protocol steps on page 1.119 - 1.122 of Molecular Cloning, Volume 1 book
Put in 37 degree shaker w/ the SOC for 1 hour. Spun down, supernatant removed to ~50ul each, resuspended, plated and spun onto correct antibiotic plates w/ LB. Put in 37 degree incubator overnight at 7:45 pm. Will take out tomorrow morning. |