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For reliability, a majority of our actual experimentation has followed protocols provided by the supplier of relevant projects, including enzymes (NEB) and kits (Qiagen). The Qiagen maxiprep kit was replaced with a Bio-Rad kit to obtain higher yields with lower processing times.

Assembly Line Flow

Bio-Brick Creation

Design Primers


Run Gel

- Check Primer concentration

- Check Primer design

Gel Extraction

Digestion with Restriction Enzymes

CIP Treatment

PCR purification


Transform and pick colonies

Miniprep and determine concentrations

Restriction Map

Retransform with chosen plasmid

Maxiprep and determine concentraions


Bio-Brick Verification: Virus Harvest

Prepare cell culture media

Seed cells

Check health of cells


Check phenotype 24hrs post transfection

Transform correct try using XL-10 cells

Maxiprep and ethanal purify the try and packaging plasmids

Seed 293FT cells

Check health of cells 24hrs later

Transform the vector and packaging plasmids using the CaPO4 method

Check for fluorescence after 24hrs

Harvest virus and concentrate using ultra-centrifugation

Please see Making A Lentivirus for protocols

Bio-Brick Verification: Quality Control

Grow cells to confluence

Seed cells

Infect cells with virus

Change media next day

Cellular experiments

- Microscopy


Please see Making A Lentivirus for protocols