Define the scope of this guide - Githubissues

hammerlab / t-cell-guide

Human Primary T cells: A Practical Guide

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Define the scope of this guide #1

Closed hammer closed 6 years ago

hammer commented 6 years ago

I would think we want to cover isolation, activation, culture (pre- and post-activation), and proliferation assays for sure. Do we want to cover electroporation of plasmids, mRNA, and RNPs? Experiences w/ specific sgRNAs and Cas9 variants?

armish commented 6 years ago

I was thinking, we would cover the basics and keep the guide simple by giving details on the things we tried and then H/Ting all the alternatives.

So,

Fundamentals: centrifuge speed, seeding density, freezing/thawing ...
Isolation: Blood -> Buffy/PBMC -> T cells
Activation
Culture (media components)
Proliferation assay
Electroporation setup for plasmid, mRNA, and RNP
Basic KO protocol (with tips on how to set it up)
Money-saving options (titration of beads, 3d printed racks, ...)

Started a very basic draft here: https://docs.google.com/document/d/1LIuLDXPm1L3KvsZwtrhaZtiUjZ3i3y74uN_FcEM-gqk/edit#heading=h.z53d11cbh61v

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