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 is less sensitive/ must proceed...
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
...
assay development
E. coli Electroporation-v1-PDF
Overview
This a protocol for preparing and handling E. coli cells for electroporation. This protocol has mostly been adapted from the Noyes Lab protocol. A...
assay development
S liquid medium-PDF
Overview
For Bcc EPS production.
Materials
1000 mL:
1 g Yeast extract
10 g Na2HPO4
3 g KH2PO4
1 g K2SO4
1 g NaCl
900 mL Water
Procedure
Dissolve...
Biologics (antibody+ADC)
10x Lambda diluent (no gelatin)-PDF
Materials
1000 mL:
60.56 g Tris base
58.0 g NaCl
10 g MgSO4
dH2O to 1000 ml
Procedure
Dissolve in 800 ml dH2O, adjust pH to 7.5...
assay development
M9 agar-PDF
Materials
500 mL
1000 mL
Bacto Agar
7.5 g
15 g
dH2O
450 ml
880 ml
10 X M9 salts + glucose
50 ml
100 ml
1 M MgSO4
1 ml
2 ml
(2 mM final)
1 M CaCl2
50 µl
0.1...