lambdaMGU2 C. elegans cDNA library (ATCC® 77366)

Organism: Caenorhabditis elegans Maupas  /  Clone Type: Library  /  Depositors: IN Maruyama

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Designations lambdaMGU2 C. elegans cDNA library
Species Caenorhabditis elegans Maupas
Depositors IN Maruyama
Applications
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

Vector information:
Vector:
lambdaMGU2
Vector type: phage
Intact vector size (kb): 41.7
Construction: lambda2690, pMGU
Features:
Insert detection:
Spi+
Marker: ampR
Replicon: lambda, pMB1, m13
Related Products
Component of: ATCC 77370
Biosafety Level 1
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

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