SEED/2009/Day 3

Morning

 * each group will miniprep 1.5ml of pSB4A5.I52001 and pSB1A3.E0050 cultures
 * elute in 30ul

Afternoon

 * spec miniprep concentrations
 * Set up one 50ul digest of pSB4A5 with EcoRI and PstI
 * 5ul NEB2
 * 0.5ul BSA
 * 2.5ug pSB4A5 miniprep
 * 1ul EcoRI
 * 1ul PstI
 * leave in 37C incubator overnight

Quiz

 * Restriction digest and gel electrophoresis

Lecture

 * What are you going to learn and do in this class?
 * Cloning Process Overview
 * Characterization

Instructor Preparation

 * grow up cultures of pSB4A5.I52001 (destination vector) and 1A3.E0050 (PCR template)
 * prepare minipreps in case student prep concentrations are too low

Instructor Post-prep

 * heat kill and store miniprep digests in freezer