927
edits
E. coli transformation: Difference between revisions
From London Hackspace Wiki
no edit summary
Mycoplasma (talk | contribs) No edit summary |
Mycoplasma (talk | contribs) No edit summary |
||
| Line 21: | Line 21: | ||
* Took 1 colony from PUC19 dh5a plate and mixed with 250ul RO water | * Took 1 colony from PUC19 dh5a plate and mixed with 250ul RO water | ||
* Incubated at 98C for 15 minutes, then hold at 10C (boiling program in PCR machine) - to lyse the cells | * Incubated at 98C for 15 minutes, then hold at 10C (boiling program in PCR machine) - to lyse the cells | ||
* Put on 2 PCRs - one with M13 primers (for PUC19), one with S16 primers (for any bacteria), then hold for 4C for 20 hours. | * Put on 2 PCRs on Col program (53C annealing) - one with M13 primers (for PUC19), one with S16 primers (for any bacteria), then hold for 4C for 20 hours. | ||
* From top - ladder, M13, S16 | * From top - ladder, M13, S16 | ||
=== | === 28 Apr === | ||
* Ran on a gel. Pic to come. | |||
* Expect M13 product of about 120bp | |||
= GFP transformation = | = GFP transformation = | ||
