pGEM®-T Vector Systems
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
Catalog Number:
Size
Add competent cells
Catalog Number: A3600
Catalog Number: A3610
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.
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
- 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.
- Robles, J. and Doers, M. (1994) Promega Notes 45, 19–20.
- Clark, J.M. (1988) Nucl. Acids Res. 16, 9677–86.
- Newton, C.R. and Graham, A. (1994) In: PCR, BIOS Scientific Publishers, Ltd., Oxford, UK, 13.
Protocols
Specifications
Catalog Number:
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 |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
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 |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for detailed storage recommendations.
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