| Products | GPS -M Mutagenesis System from New England Biolabs |
| Company | New England Biolabs |
| Item | GPS -M Mutagenesis System |
| Price | $300.00 |
| Description | The GPS-M Mutagenesis System enables you to insert a transposable element of your own design into your target DNA by in vitro transposition. It uses TnsABC* Transposase to insert a Tn7-based transposon randomly into the DNA target. Target DNA may be a plasmid, cosmid, BAC or purified chromosomal DNA, depending on your interests (6-8). If the insertion site is within a translated gene segment, this will normally result in a null (loss of function) mutation. There is minimal site preference for insertion, so disruption of any open reading frame should be possible. Due to target immunity, only one insertion occurs per DNA molecule. Target immunity acts in vivo over a distance of ~190 kb.Therefore, the in vitro reaction produces a population of target DNA molecules each containing the transposable element at a different position. To facilitate adaptation of the transposable element to your particular mutagenesis application, the transposon donor (pGPS3) can be customized by adding to or replacing the kanamycin resistance marker in the Transprimer. For ease of manipulation by the user, the donor plasmid can be grown easily in standard laboratory E. coli strains; the vector backbone carries Ampr and a pUC origin of replication. To destroy unreacted donor molecules and avoid undesirable reaction products, the donor can be destroyed by digestion with the rare-cutting enzyme PI-Sce I (VDE). For applications in which the mutagenized DNA will be transformed into naturally-competent organisms (which take up single DNA strands), a protocol is supplied for filling-in and ligating the gaps left by the insertion transposition reaction. Notes: The Kanr marker on the Transprimer is not designed to express in Gram positive organisms or eukaryotic cells. Transposon ends have stop codons in all frames, so translational fusions are not practical. |
| Info | New England Biolabs32 Tozer Rd. Beverly, MA 01915-5599 Customer Service: 800-632-5227 Tech Support: 800-632-7799 Fax Number: 978-921-1350 Web Site: http://www.neb.com |