What is the pGEM t Easy Vector?

The pGEM®-T Easy 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 a TOPO vector?

TOPO cloning is a molecular biology technique in which DNA fragments are cloned into specific vectors without the requirement for DNA ligases. For “blunt end” TOPO cloning, the recipient vector does not have overhangs and blunt-ended DNA fragments can be cloned.

How much DNA is needed to TOPO clone?

For transformation you should not use more than 50-100ng total DNA. Before TOPO cloning, you should purify the PCR product from enzymes, buffers and primers (spin column or PEG precipitation).

What’s the difference between pGem-T and easy vector?

The only difference between pGEM-T and pGEM-T Easy is in the multiple cloning site (MCS). The MCS of the pGEM-T Easy Vector contains sequences on either side of the insert that are recognized by the restriction enzymes Not I and EcoR I.

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. For Research Use Only.

What is the pGem-T sequence and map parental vector?

pGEM-T Sequence and Map Parental vector for TA cloning of PCR products. The insertion site is flanked by BstZI sites. This vector is also known as pGEM®-5Zf(+).

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.