What is a pGEM t plasmid Vector?

What is a pGEM t plasmid Vector?

The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments.

What is pGEMt?

pGEM-T Vector Information Description The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. The coding sequence was inserted by TA cloning. Most commercially available competent cells are appropriate for the plasmid, e.g. TOP10, DH5α and TOP10F´, JM109.

What is A TA cloning vector?

TA cloning (also known as rapid cloning or T cloning) is a subcloning technique that avoids the use of restriction enzymes and is easier and quicker than traditional subcloning. Such PCR amplified inserts are cloned into linearized vectors that have complementary 3′ thymine overhangs.

Which enzyme is used in TA cloning?

Taq polymerase
The TA cloning method takes advantage of the terminal transferase activity of some DNA polymerases such as Taq polymerase. This enzyme adds a single, 3′-A overhang to each end of the PCR product.

What are the T-overhangs for easy PCR cloning?

T-Overhangs for Easy PCR Cloning: The pGEM ®-T and pGEM -T Easy Vectors are linearized vectors with a single 3´-terminal thymidine at both ends. The T-overhangs at the insertion site greatly improve the efficiency of ligation of PCR products by preventing recircularization of the vector and providing a compatible overhang for PCR products

How long does it take to ligate a pGem-T vector?

Alternatively, a double-digestion may be used to release the insert from either vector. Rapid Ligation:The pGEM®-T and pGEM -T Easy Vector Systems are supplied with 2X Rapid Ligation Buffer. Ligation reactions using this buffer may be incubated for 1 hour at room temperature.

What are the recognition sites for the pGem-T vector system?

The pGEM®-T Easy Vector multiple cloning region is flanked by recognition sites for the restriction enzymes EcoRI, BstZI and NotI, providing three single-enzyme digestions for release of the insert. The pGEM®-T Vector cloning region is flanked by recognition sites for the enzyme BstZI.

Is there a GenBank accession number for the pGem-T easy vector?

There is no GenBank® Accession Number for the pGEM®-T Easy Vector. The parent vector is linearized at the position indicated by * in this pGEM®-T Easy Vector Sequence and a “T” is added at each end. What’s in the box?