Second Attempt: Restriction Digest of pBR322

 

Materials:

Methods:

1. thaw pBR322 in vector form
2. set up a two digests for pBR322 cut with EcoRI in a 1.5ml eppendorf tube, by first pipetting the following into two seperate tubes:

   

3. perform two dilutions of EcoRI: and and pipet each into separate tubes (here diluted with   and pipetted , but can also try pipetting instead of diluting)
4. mix the contents well by gently tapping the bottom of the tube with finger (avoid bubbles). EcoRI cuts pBR322 at one site.
5. briefly spin down the contents of the tube in centrifuge.
6. incubate for 1.0 - 1.5 hours
7. inactivate the restriction enzyme with 15 min

Results: ( 1/29/99)
Performed 2 digests as specified above.