BioSample

A human lens library (by) was normalized by self-subtraction. One portion of double stranded plasmid DNA representing the library was linearized by NotI. This NotI digested library was used as a template for biotinylated RNA synthesis using SP6 RNA polymerase. Another portion of the double stranded plasmid library was converted to single-stranded circles in vitro using Gene II and Exonuclease III (Life Technologies). Single-stranded DNA (1 mg) was hybridized (C0t 500) with 41 mg of Bio-RNA and vector blocking oligonucleotides. The hybridized Bio-RNA/ss-circles were removed by streptavidin:phenol extraction. EST analysis was performed on the library at the NIH Intramural Sequencing Center(NISC).