Sample Solution

The pBluescript phagemids refers to plasmids having phage origin. It is the best serving cloning vector which is designed towards simplifying the most commonly utilized cloning as well as sequencing protocols, that included the nested deletions construction towards DNA sequencing, as well as RNA transcripts generation of both in vitro as well as site-specific mutagenesis featuring gene mapping. The pBluescript has extensive polylinker containing 21 of unique restriction enzyme recognition sites. the polylinker are Flanked with T7 as well as T3 RNA polymerase promoters which are used towards in vitro RNA synthesis. The promoter’s choice serves towards initiating transcription which indeed determines the strand to be cloned by insertion into the polylinker that will be indeed transcribed.

The transcripts were isolated from both the tyrosine kinase inhibitor treated as well as the untreated KCL22 cells. For successful cloning, RNA was extracted subsequently cDNA was subjected to amplification using PCR. The plasmids were purified, digested and the amplified cDNA was cloned and transformed into vector. Human leukemic cell line (KCL22) were selected for this study.

The Kcl22 cell line is being used extensively owing to its use as exceedingly sensitive target in vitro for various assays. Cultures of ATCC stock exhibit sensitivity towards assessment of human natural killer activity. They are Chronic myelogenous leukemia, BCR-ABL1 positive cell line.These are hematopoietic stem cells having capacity to differentiate to form erythroid, megakaryocytic lineages. The entire RNA content was extracted from KCL22 cell lines, and it was subjected to reverse transcription. T