E. coli transformation: Difference between revisions
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= E coli transformed with PUC19 using ampicillin selection and electroporation = | = E coli transformed with PUC19 using ampicillin selection and electroporation = | ||
See [ | See [https://docs.google.com/document/d/1x5DkcQTWwBIKxbLGHKAjH2Sw1wdQL2B12tLCeF21b1c/edit?usp=sharing here] for protocol, successfully used by Tom H. | ||
=== 28 mar 15 === | === 28 mar 15 === | ||
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* Plate onto LBA. | * Plate onto LBA. | ||
* No e coli survived on plates, whether with Amp or not. Could be various reasons. | * No e coli survived on plates, whether with Amp or not. Could be various reasons. | ||
=== 11 Apr 15 === | |||
* Following main gdocs protocol above. Initial 2x10ml cultures started off at 13:00. | |||
= GFP transformation = | = GFP transformation = | ||
Revision as of 12:09, 11 April 2015
E coli transformed with PUC19 using ampicillin selection and electroporation
See here for protocol, successfully used by Tom H.
28 mar 15
- 2ml ecoli culture (bottle on shelf) incubated in 200ml LB (bottle in fridge)
- -ve control is untransformed ecoli from same culture put on LBA
- +ve control is transformed ecoli from same culture put on LB plates
- Transform ecoli with PUC19 using electroporation
- Plate onto LBA.
- No e coli survived on plates, whether with Amp or not. Could be various reasons.
11 Apr 15
- Following main gdocs protocol above. Initial 2x10ml cultures started off at 13:00.
GFP transformation
Growing chemically competent E. coli
Here is the protocol we used with UCL to grow competent cells during the making of the public biobrick. So we need to get E. coli (one possibility. Could also try NCBE. They sell e. coli as a transformer kit replacement part. Finally these are suggested for schools by NCBE: Sciento, Blades Biological and Philip Harris Education), LB - £66, 0.1M CaCl2 - £32.50, M9 salts - £57.60, MgSO4 - £20, bacteriological agar solution - £42.90, thiamine - ~£20, D glucose - ~£20, and plates. All links are only the first ones I found, maybe cheaper options available.
