Team:UCL/Project/Protocols

(Difference between revisions)
 Revision as of 11:29, 25 September 2013 (view source)← Older edit Revision as of 11:45, 25 September 2013 (view source)Newer edit → Line 58: Line 58:

- This protocol is for the stable transfection of eukaryotic adherent cell types in a 6-well plate, with plasmid DNA that ought to be of as high a purity as possible. + This protocol is for the stable transfection of eukaryotic adherent cell types in a single well of a 6-well plate, with plasmid DNA that ought to be of as high a purity as possible. When transfecting multiple wells, make a 'master mix' with 10% of

Line 64: Line 64:

- 2) Incubate cells in their normal growth conditions (37^0 C and 5% CO2). + 2) Incubate cells in their normal growth conditions (37^0 C and 5% CO2)for 24 hours.

- 3) + 3) Dilute 2µg of DNA dissolved in TE buffer (min conc. 0.1µg/µl) with serum, protein and antibiotic free medium (to avoid macromolecular interference with complex formation) to a total of 100µl. Mix. +

+

Revision as of 11:45, 25 September 2013

There are two solutions to $$ax^2 + bx + c = 0$$ and they are $$x = {-b \pm \sqrt{b^2-4ac} \over 2a}.$$

Bacterial Lab Protocols

This protocol is for the stable transfection of eukaryotic adherent cell types in a single well of a 6-well plate, with plasmid DNA that ought to be of as high a purity as possible. When transfecting multiple wells, make a 'master mix' with 10% of

1) The day before transfection, seed 0.9-4x10^5 cell per well of the six well plate with 2ml of appropriate growth medium. This should produce a confluence of 40-80% for the next day's transfection.

2) Incubate cells in their normal growth conditions (37^0 C and 5% CO2)for 24 hours.

3) Dilute 2µg of DNA dissolved in TE buffer (min conc. 0.1µg/µl) with serum, protein and antibiotic free medium (to avoid macromolecular interference with complex formation) to a total of 100µl. Mix.