The deposited library was preserved by mixing one volume with three volumes of SM buffer and freezing in liquid nitrogen.
Top agarose of 0.7% agarose in NZYM is recommended.
Vector
Tissue: thymus, female, Caucasian, 23 yr
DNA: cDNA
Digest: Complete
Insert size range: 0.4 - 2 kb
Genome Equivalent: 0.0
Coverage: Clones for "complete" coverage: 45000.0
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.
Comments
The original cDNA was prepared by random hexanucleotide priming of poly(A)+ RNA. It was amplified by PCR, annealed to a Cot value of 59.0 mol.sec/liter, re-amplified by PCR, and digested with EcoRI before insertion into the vector.
The deposited library was preserved by mixing one volume with three volumes of SM buffer and freezing in liquid nitrogen. The reported titer was determined on Escherichia coli C600 (ATCC 47048) after thawing a frozen sample.
IMPORTANT. This is a normalized library. Do not amplify. The library has already been amplified once and further amplifications may decrease the normalization efficiency due to bias in the biological selection.
The number of independent recombinants refers to the original cDNA library. Coverage of the normalized library was calculated from the number of clones identifed for a set of cDNA sequences.
Top agarose of 0.7% agarose in NZYM is recommended.
References
Patanjali SR, et al. Construction of a uniform-abundance (normalized) cDNA library [published erratum appears in Proc. Natl. Acad. Sci. USA 88: 5066, 1991]. Proc. Natl. Acad. Sci. USA 88: 1943-1947, 1991. PubMed: 1705712
