PCR-TRAP Products PCR-TRAP Cloning System - for cloning PCR products PCR-TRAP Vector Primer Kit - for cloning PCR products Riboprobe Primer Set - for creating Riboprobe templates AidSeq Primer Set A - for sequencing with T7 and SP36 primers AidSeq Primer Set B - for sequencing with M13 primers AidSeq Primer Set C - for sequencing with Lseq and Rseq primers For positive-selection cloning of PCR products through blunt-end ligation. Allows direct cloning of PCR products without purification. GenHunter has developed a highly efficient blunt-end PCR-TRAP Cloning System. This system uses a third generation cloning vector that features positive selection for cloning PCR products (see figures). Only recombinant plasmids confer antibiotic resistance, making PCR-TRAP extremely efficient. There is no need for any post-PCR manipulation before cloning, since a significant fraction of PCR products do not contain the 3' overhanging A (Clark, 1988, Nucleic Acids Res. 16:9677) The mechanism of this unique cloning system involves the phage Lambda repressor gene, cI, which has been incorporated into the PCR-TRAP Vector. When transcribed, the gene codes for a repressor protein which binds to the Lambda right operators Or1 to Or3 of the cro gene, turning off the promoter which drives the TetR gene on the plasmid. Therefore, cloning the PCR product directly into the cI gene leads to the inactivation of the repressor gene and thus the expression of the TetR gene. So the PCR-TRAP Cloning System is indeed a cloning TRAP! The cloned PCR product can be quickly and easily retrieved by colony-PCR for the purpose of insert confirmation or probe generation using primers which flank the cloning site (provided in the kit). If cDNA amplified by an RNAimage or RNAspectra� Kit is cloned into a PCR-TRAP System, the insert can also be excised by HindIII digestion. The flanking primers also allow the cloned PCR product to be readily sequenced. The kit contains GH Competent cells as well as a positive control, the lacZ gene, which after PCR and cloning will confer blue color upon X-gal staining. This kit is also ideal for conducting reverse Northern blot screening for positive clones during differential display (Ref. 2 and 3). For a Partial Map of the PCR-TRAP Vector click here. Refernces: Liang, P. et al.: Differential display using one-base anchored oligo-dT primers. Nucleic Acids Res. 1994, 22:5763-5764. Ikeda, S. et al.: An aquaporin-like gene required for the Brassica self-incompatibility response. Science. 1997, 276:1564-1566. Taylor, G.A. et al: Identification of a Novel GTPase, the Inducibly Expressed GTPase, That Accumulates in Response to Interferon [gamma]. J. Biol. Chem. 1996, 271:20399-20405. More Publications By GenHunter Customers   Cat. No. PCR-TRAP Cloning System P404 Detailed protocol for cloning included. Shipped on dry ice via overnight delivery. PCR-TRAP cloning system. A schematic diagram of the PCR-TRAP cloning vector before and after cloning of a PCR product. PCR-TRAP Cloning Vector ligated with a 450 bp PCR amplified LacZ gene was transformed into E. coli and plated on a tetracycline plate. Blue colonies are positive for insert, while white colonies indicate background colonies. Colony PCR analysis of the 450 bp LacZ inserts cloned into the PCR-TRAP vector. Lane one is a 450 bp LacZ PCR fragment before cloning. Lane 2 is a 100 bp DNA ladder as a MW standard. Lanes 3-10 are randomly picked blue TetR colonies that were checked for insert by colony PCR using a pair of primers located 60 bp away from either side of the cloning site. All positive clones will give a PCR product 120 bp bigger than the actual insert before cloning. The flanking primers can then be used for sequencing. Note all clones checked contain the expected insert. This kit contains some of the essentail components of the PCR-TRAP Cloning System. Other components such as T4 DNA ligase, Component cells, and Tetracycline are not provided.   Cat. No. PCR-TRAP Cloning Vector Primer Kit P405 Individual components for the PCR-TRAP Cloning System sold separately Description Cat. No. Volume PCR-TRAP Cloning Vector V101 60 �L T4 DNA Ligase (200 units/�l) L101 30 �L Colony Lysis Buffer L102 5 �L 10X Ligation Buffer L103 70 �L Lgh Primer (2 �M) L201 250 �L Rgh Primer (2 �M) L202 250 �L Tetracycline (20 mg/mL) L401 1 �L GH Competent Cells (6 vials) L301 3 �L Lseq Primer (2 �M) L402 100 �L Rseq Primer (2 �M) L403 100 �L

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