Blackbourn laboratory methods

2

• 293 cell culture medium
• 2-dimensional gel electrophoresis

A

• Alkaline lysis of miniprep of plasmid DNA
• Antibody depletion

B

• BACK50, batch testing of
• BACK50, luciferase testing of
• BACK50, production of
• BAC recombination protocol using galK
• Bac-to-Bac Baculovirus Expression System (Invitrogen)
• BCBL-1 cells, reactivation of
• B cell transfection by electroporation
• Buffer E for cell lysis (Western blotting)

C

• Calibrating the CO2 incubator
• CAT reporter assay
• Citrate antigen retrieval buffer
• Complement attack assay
• Complement attack assay (revised)
• Coomassie Blue staining of SDS-PAGE gels
• Culture of tDMVEC

D

• DNA loading buffer
• DNA probes, labelling for Southern and Northern blotting
• DNAse treatment
• Dual luciferase assay

E

• Electrophoretic mobility shift assay (EMSA)
• Endothelial cell medium
• Enhanced chemiluminescence (ECL) reagents
• Enzyme-linked immunosorbent assay (ELISA)

F

• FACScalibur
• Flow cytometry
• Flow cytometry for vOX2-EGFP and CD200_EGFP expressing SVK cells

G

• G418 sensitivity of SLK cells
• Gel Electrophoresis

H

• Haematoxylin and eosin (H&E) staining of tissues section
• Harvesting cells for luciferase assay
• Harvesting dishes of tDMVEC cells
• HeLa transfections
• Homemade chemically competent E.coli
• Homemade chemically competent E.coli, transformation of plasmid into
• Hybridisation of Hybond nylon membrane (Amersham)

I

• IFA acetone/methanol fixation
• Immunocytochemistry
• Immunoprecipitation
• In-gel digestion of Coomassie-stained protein
• Intracellular IFN assay

K

• KSHV, production of
• KSHV, qPCR for copy number
• KSHV, qPCR for copy number (absolute quantification)

L

• Labscan
• Lab stocks instructions
• Lentivirus for transduction, production of

M

• Membrane protein purification
• MTT colorimetric assay
• Mycoplasma testing of adherent cells

N

• Northern blotting

O

• OLIGO 6 instructions
• ORF4 immunoprecipitation

P

• PBMC preps for buffying T cell clones
• PCR primer dissolution
• Permanent plasmid stocks
• Phenol extraction
• Plasmid Purification
• Plasmids on filter paper
• PolyA mRNA by oligo(dT) bulk preparation
• Preparation of EMSA probes
• Prepartation of competent JM109 E.coli cells
• Proteinase K treatment of PCR products for cloning

R

• Retrovirus for transduction, production of
• RNA preparation with Trizol
• RTA/BACK50 batch testing

S

• SDS-PAGE gels
• SDS-PAGE loading buffer
• SF9 insect cells, growth of
• Shipping instructions for sera
• Shipping instructions for virus
• Silver staining of gels
• STM cell lysis

T

• TAE buffer, 50x
• TaqMan
• T cell clone culture medium
• T cell clonning with antigen specific tetramers
• T cells, IFN assay
• T cells, staining for CD200R
• Total RNA purification
• Transformation of competent cells
• TSA simple amplification
• Tumour infiltrating lymphocyte preparation methods

W

• Western blotting
• Western blotting (for dummies)