pAL-F (ATCC® 87563)

Applications: vector containing primer sites useful for sequencingvector for generating nested deletions of cloned insertsvector permitting visual detection of recombinants  /  Depositors: A Ahmed, L Podemski

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Designations pAL-F
Depositors A Ahmed, L Podemski
Biosafety Level 1
Vector Information
Size (kb): 8.8400001525878910
Vector: pAL-F (cosmid)
Marker(s):ampR,kanR,strS
Construct size (kb): 8.84000015258789
Features: insert detection: lacZalpha
marker(s): ampR
marker(s): kanR
marker(s): strS
other: IS1 transposable element
other: lambda cos site
replicon: pMB1
replicon: rop (copy number control)
MCS: EcoRI...HindIII
coding sequence: galK
Applications
vector containing primer sites useful for sequencing
vector for generating nested deletions of cloned inserts
vector permitting visual detection of recombinants
Comments
Restriction digests of the vector give the following sizes (kb): HindIII--8.8; BamHI--8.8; EcoRI--8.8.
To obtain ordered deletions of a genomic fragment, the DNA of interest is cloned into the BamHI site, followed by digestion of the plasmid with one of 4 restriction enzymes (KpnI, SalI, XhoI, SacI) and subsequent religation to obtain a set of subclones.
IS1-promoted deletions can be selected by plating each subclone on deoxygalactose-containing media. Appropriate deletion clones should be Gal- and strS.
Cloning vector useful for sequencing genomic DNA inserts by generation of an ordered set of deletion subclones.
Sequencing of the cloned inserts can be performed using the -23 IS1 primer (5' AAA ACA CCA TCA TAC ACT AA 3') published in Methods in Enzymology 155:177-204, 1987.
Media ATCC Medium 1065 (see below) plus ampicillin (100 ug/ml) plus kanamycin (50 ug/ml) ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0 L
Growth Conditions
Temperature: 37.0°C
References

Ahmed A, Podemski L. Use of ordered deletions in genome sequencing. Gene 197: 367-373, 1997. PubMed: 9332387

Shipped frozen
Shipping Information

 Frozen glycerol stock of Escherichia coli containing the cosmid.

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