zhourui
Diagnostics
Pouring LB plates from prepped media-PDF
Notes
Melt LB ahead of time because it cools to 55 degrees slowly. A good way to do this is melt it in the...
Biologics (antibody+ADC)
Nested RT-PCR-PDF
Polymerase dependent expansion of a DNA amplicon
Korey Griffin Resource List
qPCR Advantages
Collect reliable data at low cycle numbers. Traditional PCR...
Biologics (antibody+ADC)
RNA extraction-PDF
Comparison of purification approaches
Guanidine isothiocyanate extraction coupled with lithium chloride precipitation
Lyses cells, extracts cellular RNA and denatures proteins all at once in many...
assay development
Supplemented M9 broth (M9sup)-PDF
Materials
50 mL
500 mL
1000 mL
dH2O
43 ml
430 ml
860 ml
10 X M9 salts + glucose
5 ml
50 ml
100 ml
10% Casamino acids
1 ml
10 ml
20 ml
1% Thiamine
50 µl
0.5 ml
1 ml
1...
assay development
Making frozen permanents-PDF
Materials
Cryo vials
Overnight bacterial culture
LB (50% glycerol)
Procedure
Pipette 400 μL LB 50% glycerol and 600 μL overnight culture into a cryo vial. Vortex,...
assay development
Digestion-PDF
Overview
Standard DNA single or double digest of variable volume V using NEB restriction enzymes.
Materials
X μL DNA template
10X NEB Digestion Buffer Y (Y...
assay development
Electrocompetent cells-PDF
Materials
GYT (glycerol, yeast extract, tryptone)
• 10%(v/v) glycerol
• 0.125% (w/v) yeast extract
• 0.25% (w/v) tryptone
DI water
10% Glycerol
Special Equipment
Centrifuge
...
assay development
Chung cells-PDF
Materials
Overnight E. coli culture
50 mL Falcon tubes
LB
TSS
1.5 mL centrifuge tubes
Storage:
Freezer box
Transformation:
LB or TSS with 20 mM glucose
...
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