To prepare phagemid from lambdaMGU2, grow recombinants on a RecA- host expressing the Cre protein (E. coli 1046[pCRE1], ATCC 77368) and select for ampicillin resistance.
Strain of C. elegans used for constructing this library is C. elegans N2 which is a wild type strain.
Vector
Library information: Genome:Caenorhabditis elegans Strain: N2 Type of insert: cDNA Vector: lambdaMGU2 Insert size range (kb): 2-3 kb average Vector ends: Modification: BamHI Number of independent recombinants: 1.0 x 107 Titer: 5 x 107 pfu/mL
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
Frozen bacteria-free phage lysate
Comments
Strain of C. elegans used for constructing this library is C. elegans N2 which is a wild type strain.
Single-stranded DNA may be recovered from phagemid constructs using M13KO7 helper phages.
To prepare phagemid from lambdaMGU2, grow recombinants on a RecA- host expressing the Cre protein (E. coli 1046[pCRE1], ATCC 77368) and select for ampicillin resistance. The pMGU product is 4.185 kb.
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 - 3'gam/BamHI/5'gam - XhoI - loxP - SalI - lambda N.
Efficiency of phagemid recovery is approximately 20%. Plasmid pCRE1 may be a low level contaminant, but is easily distinguished from pMGU DNA.
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
