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DNA ISOLATION AND PLASMID PURIFICATION

> Phenol extraction and ethanol precipitation.

   

> Large Scale Plasmid Prep for 100 ML Superbroth Cultures

   

> Plasmid Miniscreen Protocol (Boil Prep)

   

> Alkaline Lysis Miniprep

   

> Alkaline Lysis Miniprep Modified 1

   

> Alkaline Lysis Miniprep Modified 2

   

> Amplification of plasmids with chloramphenicol

   

> Single-Strand DNA Prep. Large Scale

    > Titre of Bacteriophage
  > Large Scale Bacteriophage Prep

 

LARGE SCALE BACTERIOPHAGE PREP

1. Inoculate 500mL of LB with 100ul of SURE cells. Grow at 37ºC with agitation for several hours.

2. Pick bacteriophage plaque from a plate and inoculate into 1mL of LB. Let stand at room temp. For 1-2 hours.

3. Inoculate bacterial culture with 1 mL of phage suspension. Grow at 37ºC with agitation for 5-6 hours.

4. Spin down the suspension at 12,000g for 10 min.

5. Withdraw the supernatant( make sure not to disturb the bacterial pellet).

6. Add a few drops of Carbon Tetrachloride to the supernatant and store at 4ºC.

7. See TITRE OF BACTERIOPHAGE to titre phage.