Creation+of+L4440

**Welcome to the Creation of the Cloning Vector Page!**

__Background__:

__Procedure__: 1. 2 cultures needed to make mini-preps:
 * LB tubes
 * AMP (1,000 x) 1 uL/mL
 * ~500 uL L4440 (in DH5alpha) per culture

2. Place the cultures in a shaking water bath for 24 hours.

3. Conduct 4 Zippy mini-preps to separate the L4440 plasmid from the //E. coli// DH5alpha strain host cells.

4. Digest the L4440 vector using the following protocols.
 * 1 ul of each enzyme (HindIII, SacII)
 * 10 ul L4440 vector
 * 3 ul 10x buffer
 * 15 ul dd water

5. Put in a 37 degrees Celsius water bath.