Gene cloning and vector construction are some of the most used techniques in molecular biology. Bioline has an extensive range of cloning reagents designated to prepare your targets. Short DNA fragments or even whole genes are inserted into plasmids or virus chromosomes ligation of the fragments/inserts can be performed using a T4 DNA ligase. Bioline developed a quick-stick T4 DNA ligase which together with an optimized buffer improves enzyme activity thus increasing the ligation of your insert into the vector.
The vectors with ligated insert are then transformed into a suitable host which is more commonly referred to as competent cells. After cultivation the hosts are screened for cells that contain the recombinant plasmid or vector. This is commonly done by adding two selection tools. An antibiotic resistance gene which allows only transformed cells to replicate and mechanism to indicate positive ligation, commonly done by blue white colony screening. Bioline provides a range of competent cells to suit your specific application. All competent cell lines from Bioline contain the Lac-Z operon to perform blue/white screening with X-gal-IPTG.
These cells containing the recombinant plasmid or vector can then be isolated allowing for downstream applications like sequencing the gene of interest, or study of gene expression/protein production.