Sample Solution

quality was affirmed by real time qPCR. Amplified product of TROVE2 transcript was cloned into vector subsequent to digestion with selected restriction digestion with pst1 and sgf1. Then, it was ligated by using Bioline QS ligase enzyme. Transformation was carried by heat shock method. Blue white selection strategy was employed to select the transformed colonies. Colony PCR was performed for ascertaining the proper cloning. Agarose gel electrophoresis was employed to separate DNA fragments and analysed fragments cross checked for ascertainment.

Results:

Tissue culture of KCL22 cells were grown as per routine standard tissue culture conditions. Treated KCL22 cellular samples were grown with media being supplemented with tyrosine kinase inhibitor( TKI) drug for a period of 72h whereas untreated cells were indeed cultured in medium devoid of TKI drug. The morphology of KCL22 cells were subsequently observed by inverted microscope. Viability as well as distribution of size was observed using automated cell counter of BioRad T make.

The average percentage has relatively dropped with treated cells (Table 1A and B).

Table 1A and B:

Preparation of plasmid from E.coli: Preparation of pBluescript SK+ plasmid in E.coli was carried over by growing the E.coli cells with LB media supplemented with ampicillin antibiotic. pBluescript SK+ plasmids were purified using ‘plasmid purification’ spin column kit. The concentration of purified plasmid was ascertained using Invitrogen Qubit flourometer by using Quant-IT dsDNA BR dye. Results indicate that the fractions were pure.