Our website does not fully support your browser.

We've detected that you are using an older version of Internet Explorer. Your commerce experience may be limited. Please update your browser to Internet Explorer 11 or above.

We believe this site might serve you best:

United States

United States

Language: English

Promega's Cookie Policy

Our website uses functional cookies that do not collect any personal information or track your browsing activity. When you select your country, you agree that we can place these functional cookies on your device.

PCR Cloning

T-vector cloning, or TA cloning, is a convenient method for cloning PCR products generated with Taq DNA Polymerase. The pGEM®-T vectors are a popular choice for general PCR cloning. These vectors are ready to use in ligation reactions; prepared by cutting with a restriction endonuclease that creates a blunt end and adding a 3´ terminal thymidine (T) to both ends. The 3´ T-overhangs at the insertion site greatly improve ligation efficiency of a PCR product into the plasmid by preventing recircularization of the vector and providing a compatible overhang for PCR products with 5' A-overhangs. The pGEM®-T Easy Vector systems come with competent cells included.

The pTARGET™ Vector is used for cloning and expression of PCR products in mammalian cells. Then, for sequencing inserts cloned into this vector, the pTargeT™ Sequencing Primer is available.

Participate in Promega's 31 Days of Discovery to win fun prizes this holiday season! Sign Up ›

Filter By


Vector Type

Promoter

PCR Accessories

Shop all PCR Cloning products

Showing 4 of 4 Products

An Introduction to PCR Cloning

PCR products generated using a non-proofreading DNA polymerase which lacks 3’ to 5’ exonuclease activity, such as Taq DNA polymerase, are left with a single template-independent nucleotide, deoxyadenosine (A), at the 3´ end of the amplified fragments. This single-nucleotide overhang allows hybridization with and cloning into vectors that have a complementary 3´ single deoxythymidine (T) overhang.  

T vectors are linearized plasmids that have been treated to add T overhangs to match the A overhangs of the PCR product. PCR fragments that contain an A overhang can be directly ligated to these T-tailed plasmid vectors with no need for further enzymatic treatment other than the action of T4 DNA ligase.

The high-copy-number pGEM®-T and pGEM®-T Easy Vectors contain T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the coding region for the a-peptide of ß-galactosidase, allowing blue/white selection of recombinant clones.

Both the pGEM®-T and pGEM®-T Easy Vectors contain numerous restriction sites within the multiple cloning region. 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. Alternatively, a double digestion may be used to release the insert from either vector.