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pGEM®-T Vector Systems

pGEM T Easy Vector System 20 reactions
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Convenience for Cloning PCR Products

  • Insert excision with a BstZI single digest
  • Ligation can be completed in 1 hour at room temperature
  • Available with or without competent cells

Size

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Catalog number selected: A3600

$ 156.00
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pGEM®-T Vector Systems
20 reactions
$ 156.00
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Efficient T-Vector Cloning with Blue/White Selection

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. The pGEM®-T Vector is ready to use in ligation reactions, prepared by cutting the pGEM®-5Zf(+) Vector with EcoRV and adding a 3´ terminal thymidine to both ends. These single 3´-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmid by preventing recircularization of the vector and providing a compatible overhang for ligation of PCR products with A overhangs.

pGEM®-T Vector Map and Sequence

pGEM-T Vector.

The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. X65308). The pGEM®-T Vector was created by linearizing the pGEM®-5Zf(+) Vector with EcoRV at base 51 and adding a T to both 3´-ends. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt).

In the pGEM®-T Vector, T7 and SP6 RNA polymerase promoters flank a multiple cloning region within the α-peptide coding region for β-galactosidase. Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication.

The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Alternatively, a double digestion may be used to release the insert from the vector.

The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components.

References

  1. Mezei, L.M. and Storts, D.R. (1994) In: PCR Technology: Current Innovations, Griffin, H.G. and Griffin, A.M., eds., CRC Press, Boca Raton, FL, 21.
  2. Robles, J. and Doers, M. (1994) Promega Notes 45, 19–20.
  3. Clark, J.M. (1988) Nucl. Acids Res. 16, 9677–86.
  4. Newton, C.R. and Graham, A. (1994) In: PCR, BIOS Scientific Publishers, Ltd., Oxford, UK, 13.

Specifications

You are viewing: A3600 Change Configuration

What's in the box?

Item Part # Size Available Separately

pGEM®-T Vector

A362A 1 × 1.2μg

Control Insert DNA

A363A 1 × 12μl

2X Rapid Ligation Buffer

C671A 1 × 200μl View Product

T4 DNA Ligase

M180A 1 × 100u

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

See Protocol for detailed storage recommendations.

Specifications

You are viewing: A3610 Change Configuration

What's in the box?

Item Part # Size Available Separately

pGEM®-T Vector

A362A 1 × 1.2μg

Control Insert DNA

A363A 1 × 12μl

2X Rapid Ligation Buffer

C671A 1 × 200μl View Product

JM109 Competent Cells, High Efficiency

L200A 6 × 200μl

T4 DNA Ligase

M180A 1 × 100u

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

AA

See Protocol for detailed storage recommendations.

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