Week 1: Tutorial Outline

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

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

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