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| 产品名称： | lambdaMGU2 |
|---|---|
| 商品货号： | TS214623 |
| Designations： | lambdaMGU2 |
| Depositors： | IN Maruyama |
| Applications： | Vector for constructing cDNA libraries
Vector permitting positive selection for inserts
Vector permitting production of single-stranded DNA |
| Vector： | Vector type: phage Vector size (kb): 41.7 Markers: ampR Replicons: lambda, pMB1, M13 Cloning site: BamHI Suggested host: E. coli Q358 |
| Related Products： | Component of: ATCC 77369 |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Shipping Information： | Freeze-dried bacteria-free lysate |
| Comments： | Restriction digests of the clone give the following sizes (kb): HindIII--24.1, 17.6; EcoRI--22.0, 20.0; BamHI--24.8, 16.9; XbaI--32.7, 9.0; NotI--41.7.
Efficiency of phagemid recovery is approximately 20%. Plasmid pCRE1 may be a low level contaminant, but is easily distinguished from pMGU DNA.
To prepare phagemid from lambdaMGU2, grow recombinants on a RecA- host expressing the Cre protein (E. coli 1046pCRE1, ATCC 77368) and select for ampicillin resistance. The pMGU product is 4.185 kb.
Vector useful for constructing cDNA libraries. Permits positive selection for inserts using the Spi- phenotype, and excision of phagemid by lox/cre site-specific recombination.
To enable the positive selection of inserts, the library should be plated on a P2 lysogen such as Escherichia coli Q359 (ATCC 47019).
Inserts can be amplified using the following primers flanking the BamHI cloning site: upstream 5-AAGAGGCAGAACTGGCAG-3 and downstream 5-ATCGATGCATAGCGATTC-3.
The order of the major features in the cloning region of the lambda vector is: lambda J - SmaI - SalI - loxP - EcoRI - M13 ori - ampR - pMB1 ori - HindIII - 3gam/BamHI/5gam - XhoI - loxP - SalI - lambda N. |
| References： | Maruyama IN, Brenner S. A selective lambda phage cloning vector with automatic excision of the insert in a plasmid. Gene 120: 135-141, 1992. PubMed: 1327972 |