Metabolic Engineering Class

Public group
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You asked for it and here is our summer intensive workshop to learn Metabolic Engineering and lab techniques. This is a 4 session class meeting over 4 Saturday mornings.

Price is for the full 4 class session. The class is $200 for non-members and $150 for members of BioCurious.

Session 1
Intro to bacteriology, aseptic technique
Media prep
Explain central dogma (DNA to RNA to proteins)
Describe the various types of DNA parts (promoters, orfs, etc)
Lay out experimental design: biochemistry of lycopene biosynthesis
build plasmid to combine crtE, crtB, crtI from Pantoea
How to search literature and databases
Explain PCR
How to use ApE program
How to design and order oligos

Session 2

Review experimental design and goals
Inoculate comp cell cultures
Build parts/vector by PCR
Describe origins of replication, plasmid compatibility, selection markers
Describe various cloning methods (restriction cloning, Gibson, InFusion, Golden Gate)
Gel purify parts
Clone inserts into vector (Gibson method)
Comp cell prep
Transform cells
Plate cells (explain how to streak)

Session 3
Review experimental design
Colony PCR
Describe types of sequencing (Sanger, various NGS methods)
Explain how align sequences
Run cPCR samples on gel
Discuss students' ideas for projects
Submit for sequencing

Session 4
Prepare cultures for comp cells
Review experimental design
Build contigs
Perform sequence alignment
Mini-prep verified constructs
Discuss students' projects, ethics
Transform to combine constructs

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