BioSample

Priming method: GsuI-d(T) priming ; Priming sequence: 5' -GAGCTAGTTCTGGAG(T)16VN -3' ; Directionally cloned: ; 5' cloning site: NotI site GCGGCCGC ; 5' linker/adaptor sequence: 5' -GGATCCGAC -3' ; 3' cloning site: BseRI ; Insert size range: 1.5kbp-700bp ; For PCR insert analysis: M13 forward and reverse primers ; Library is not amplified ; Recombinants (inserts): 98% ; Library complexity: 7.5 X 106 cfu ; Full-length construction (method): Ng et al, Nature Method 2005 2(2):105-111 ; The pooled tissue RNA was collected and used to construct full length enriched cDNA library and also served as template to synthesize complex first strand cDNA probe. Two high density colony arrays were made from over 110K cDNA clones and hybridized with the probes. Low abundant clones were selected by the Colony Array Subtractive Hybridization (CASH) analysis as they represented rare expressed clones. The hybridization intensities for all clones span from 0 to 300,000 counts and the low abundant selected clones in this library ranges from 0 to 13,000.