Syllabus of Jawaharlal Nehru University MSc Biotechnology - BT524 - Laboratory Techniques-II

Credits: 4

Molecular Biology

Insertion mutation of a cloned gene

Large/Medium scale isolation of an E. coli plasmid (pUC19 and pUC7 KAPA).
Estimation of amount of plasmid isolated.
Restriction mapping of the plasmid pBR322.
Isolation of a gene (neomycin phosphotransferase) from the plasmid pUC7 KAPA.
Cloning of the BamHI fragment containing the neomycin phosphotransferase gene into the BamHI site of pUC19 B/W screening.
Transformation of E. coli DH5 ( ) w ith pUC19 and also with pUC19 containing a mutated tetracycline resistance gene.
Isolation of plasmid from the selected transformants by the "miniprep" method.
Restriction analysis of the plasmid to locate position of the insert.
Expression of B-gal under different promoters T7 system and pUC19 (lac promoter), with wild type E. coli as control.

Immunology

Purification of Immunoglobulin from serum
Generation of antibody in mouse
Double diffusion
Conjugation of antibodies with Enzyme
ELISA : i) Capture ELISA
ii) Direct ELISA

Western blot
Affinity column and purification of antigen
Cell fusion for generation of Hybridoma