EQ nuclear expression vector construction experiment

First, the service introduction

Molecular clone provides a method of purifying and amplifying a particular DNA segment at a molecular level. Often contains a target gene, inserting them into a cloned carrier, forming a recombinant cloning carrier, forming a suitable host, then introducing a suitable transfer host, and then separating purified by the selected host intracellular The required cloning carrier can be obtained by inserting a DNA, thereby obtaining an amplification of the purpose
gene.
Second, Experiment Principle
The connection of foreign DNA and carrier molecules u0026 nbsp is DNA recombination, which recombined DNA is called recombinant or recombinant. The recombinant DNA molecule is an exogenous DNA molecule to be linked to an exogenous DNA molecule in the connection buffer system of Mg2 + and ATP, and a carrier molecule, respectively. DNA ligase has two: T4 phage DNA ligase and E. coli DNA ligase. Both DNA ligases have functions u0026 nbsp to connect two DNA molecules with the same viscous ends; and T4 phage DNA ligases also have an E. coli even which is not characterized, it can make two The double-stranded DNA molecule of the flat end is connected. However, the efficiency of this connection is lower than the connection efficiency of the viscous end, and can improve the connection efficiency of the flat end by increasing the T4 phage DNA ligase concentration or increasing the DNA concentration.