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== | ==Prepping Plasmid for Electroporation== | ||
===PZE-12-CFP vector from JM Kim=== | |||
* Vector Nomenclature: http://www.expressys.com/main_vectors.html | |||
===Electroporations=== | |||
{| class="wikitable" border="1" | |||
|- | |||
! ng/ul | |||
! 260/280 | |||
! 260/230 | |||
|- | |||
| 56.3 ng | |||
| 1.92 | |||
| 1.62 | |||
|} | |||
Doing 4 electroporations, in order to ensure competent cells made last week work correctly. | |||
{| class="wikitable" border="1" | |||
|- | |||
! Symbol | |||
! Media | |||
! Plasmid | |||
! Cells | |||
! Why | |||
|- | |||
| A | |||
| LA/Amp | |||
| PZE-12-CFP | |||
| My competent MG1655 | |||
| To get comp cells w/ plasmid | |||
|- | |||
| B | |||
| LA | |||
| PZE-12-CFP | |||
| My competent MG1655 | |||
| To check that my competent cells are alive | |||
|- | |||
| C | |||
| Spec/AMP | |||
| PZS4-ATTP | |||
| My competent MG1655 | |||
| To check that my competent cells transform | |||
|- | |||
| D | |||
| LA/AMP | |||
| PZE-12-CFP | |||
| Jeff's competent MG1655 | |||
| To check that my plasmid works/is correct | |||
|} | |||
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. |