Production Manuals & SOPs

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Production Manuals:

• Large Scale Growth
• Subcellular Fractions
• Preparation and Fractionation of Cell Wall Proper
• Lipoarabinomannan (LAM) and Lipomannan (LM)
• Antibodies
• Microarrays

Individual SOPs:

Media

• M001: Preparation of GAS medium
• M002: Preparation of GAS medium with tween
• M003: Preparation of Sauton's medium
• M004: Preparation of Sauton's medium with tween
• M005: Preparation of 7H9 medium
• M006: Preparation of 7H9 medium with tween
• M007: Preparation of 7H9-Dextrose medium
• M008: Preparation of 7H9-Dextrose medium with tween
• M009: Preparation of 7H11 agar plates
• M010: Preparation of 7H11-Dextrose agar plates
• M011: Preparation of Nutrient agar plates
• M012: Preparation of Complete RPMI medium
• M013: Preparation of Mycobactin HR Media and Plates

Reagents

• R001: Preparation of 32% Triton
• R002: Preparation of 1st fix for gels
• R003: Preparation of 2nd fix for gels
• R004: Preparation of Coomassie stain reagent
• R005: Preparation of Coomassie stain destain
• R006: Buffer B for polyacrylamide gels
• R007: Buffer D for polyacrylamide gels
• R008: Preparation of SDS-PAGE Tris-glycine running buffer
• R009: SDS-PAGE 10X Tricine running buffer
• R010: Preparation of western blot transfer buffer
• R011: Preparation of charring spray
• R012: Preparation of α-napthol spray
• R013: Preparation of Dittmer-Lester spray
• R014: Preparation of TAE Buffer

Native Material Production

• PP001: Establishing frozen stocks of M. tb (small scale)
• PP002: Establishing frozen stocks of M. tb (large scale)
• PP003: Large scale growth of M. tuberculosis
• PP004: Gamma-irradiation of M. tb
• PP005: Preparation of CFPs and low MW CFPs
• PP006: Preparation of CFPs using 2L Stirred Cell
• PP007: Preparation of whole cell lysate
• PP008: Preparation of subcellular fractions
• PP009: Preparation of genomic DNA
• PP010: Preparation of TX-114 proteins
• PP011: Preparation of mAGP
• PP012: Preparation of arabinagalactan
• PP013: Preparation of peptidoglycan
• PP014: Preparation of mycolic acids
• PP015: Preparation of LAM, LM, and PIM (LLP)
• PP016: Separation of LLP
• PP017: QC of LAM and LM
• PP018: Isolation of total lipid
• PP019: Purification of Con-A reactive CFPs
• PP020: Preparation of Ag85 complex
• PP021: Preparation of Ag85 A, B, and C
• PP022: Purification of MPT32
• PP023: Purification of the 16kDa ACR/HspX
• PP024: Purification of 38kDa
• PP025: Preparation of SDS soluble cell wall protein
• PP026: Preparation of PIM
• PP027: Purification of SapM
• PP028: Preparation of CFP minus LAM
• PP029: Purification of Trehalose Dimycolate (TDM)
• PP030: Purification of Trehalose Monomycolate (TMM)
• PP031: Purification of 19kDa
• PP032: Purification of Mycobactin
• PP033: Growth of M.tb for Mycobactin Production
• PP034: Preparation of Sulfolipid-1

Recombinant Material Production

• RP001: Generation of Recombinant Clone Frozen Stocks
• RP002: Establishing Stocks of Low Copy Plasmids
• RP003a: Production of Recombinant Proteins
• RP003b: Production of Recombinant Proteins Denaturing Conditions
• RP004: Production of ESAT6
• RP005: Production of CFP10
• RP006: Small- Scale Purification of Plasmid DNA
• RP007: Production of rec GlcB
• RP007msmeg: Production of rec GlcB from M. smegmatis
• RP008: Production of rec Rv1411c
• RP010: Production of rec Ag85A
• RP011: Production of rec Ag85B
• RP012: Production of rec Ag85C
• RP014: Production of rec DnaK
• RP015: Production of rec GroEL2
• RP016: Production of rec Mpt64
• RP019: Production of rec BRFB
• RP020: Production of rec Mpt51
• RP021: Production of rec HspX
• RP022: Production of rec Rv2626c
• RP023: Production of rec Rv0577
• RP025: Production of rec KatG
• RP026: Production of rec Rv3763
• RP028: Production of GroES

Antibody Production

• AB100: Cell Line Maintenance
• AB101: IgG Purification
• AB102: Antibody QC
• AB103: Monoclonal Fusion
• AB104: Screening and Subcloning
• AB105: Freezing Cell Lines

Standard Procedures

• SP003: BCA protein assay
• SP004: Lyophilization
• SP005: Operation of the savant
• SP006: Preparation of SDS-PAGE Tris-glycine gels
• SP007: Running of polyacrylamide gels
• SP008: Running of IEF strips by IPGPhor
• SP009: Running of 2D gels
• SP010: Running of 2D gels using ZOOM System
• SP011: Western blot protocol
• SP012: Silver staining
• SP013: Coomassie staining
• SP014: Quantitating and assesing purity of DNA by UV spec.
• SP015: Ligation of DNA fragments with sticky ends
• SP016: Ligation of DNA fragments with blunt ends
• SP017: Restriction endonuclease digestion of DNA
• SP018: Running of agarose gels
• SP019: Removal of SDS by paired-ion extraction
• SP020: LAL assay
• SP021: In-gel digestion of proteins
• SP022: Preparation of alditol acetate derivatives
• SP023: Operation of Orion 310 pH meter
• SP024: Operation of Corning 430 pH meter in C210
• SP025: Operation of Waters 600E HPLC system
• SP027: Operation of french press
• SP028: PCR for detection of clones
• SP029: Preparation of electro-competent M. smegmatis
• SP030: Percent SDS determination in an aqueous solution
• SP031: Operation of the N2 Bath
• SP031b: Operation of N2 Bath Meter
• SP032: Preparative Thin Layer Chromatography
• SP033: Thin Layer Chromatography
• SP034: AlamarBlue Testing
• SP035: Acid-Fast Staining
• SP036: Gram Staining
• SP037: Operation of rotovap
• SP039: ELISA
• SP040: Capture ELISA
• SP040b: IL-2 Capture ELISA
• SP042: Electroporation of M. smegmatis
• SP043: Recombinant Plasmid Map Design-VectorNTI
• SP044: Sequence Alignment-VectorNTI ContigExpress
• SP045: Operation of the HP 5890A Gas Chromatograph
• SP046: NMR of Glycolipids