1. Week 1: Tutorial Outline

2. Important Techniques:
Design and implementation of
PCR
DNA purification/gel extraction
Gel electrophoresis
Cloning
Transformation
Plasmid preps
Sequencing
Binding assays
FACS

3. To Do This Week Currently we have: Random 10mer ssDNA oligo library
Control plasmids:
OmpA + His tag, OmpA + Strep tag
Goal this week:
Plasmid library of random 10mers that is transformed into E. Coli
Begin column selection binding assay of transformed E. Coli

4. Week 1 Outline: Day 1: Day 2: Day 3: Day 4: Day 5:
PCR extension of random library
PCR purify library
Day 2:
Restriction digest of library and OmpA plasmid
Run on gel and gel extract plasmid
Ovenight ligation
Day 3:
Transformation of ligation products from Day 2 (x3) in culturing strain
Plate to determine transformation efficiency (1x)
Sequencing of OmpA control plasmids (His, Strep)
Day 4:
Count colonies to evaluate efficiency of transformation/ligation
Miniprep liquid culture transformants form Day 3
Transform into competent cells
Plate and keep in liquid culture
Day 5:
Check the transformation from Day 4 worked
Column selection
FACS