Difference between revisions of "SEED/2010/Day 3"

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(New page: == 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 pSB4...)
 
(No difference)

Latest revision as of 10:37, 2 February 2010

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