Current PDB may contain all templates for single-domain proteins according to the seminal studies in Zhang and Skolnick (2005a).
A total of 1 μg RNA was used as the template for single strand cDNA synthesis utilizing random primers and the Primescript reverse transcriptase (M-MLV, Takara, Japan).
One μg of total RNA was isolated from non-tumor colonic tissues using RNA Bee (Tel-Test, Inc., Friendwood, TX) and used as the template for single strand cDNA synthesis utilizing the Transcriptor First Strand cDNA Synthesis Kit (Roche, Indianapolis, IN) according to the manufacturer's instructions.
From 2 5 micrograms of total RNA, double-stranded cDNA was generated as a template for single-round in vitro transcription with biotin-labeled nucleotides using the Affymetrix cDNA Synthesis and In Vitro Transcription kits (Affymetrix Inc., Santa Clara, CA).
These were used to construct a single-stranded shotgun library that was used as a template for single-molecule PCR.
One microgram of total RNA served as a template for single-stranded cDNA synthesis in a reaction using oligo (dT) primers and SuperScript II (Invitrogen).
A small aliquot (~2 μl) of the resultant PCR product was then used as template for single-strand DNA (ssDNA) generation using only the N (nested) primers for all tags.
For the RT PCR, the total volume (10 μl) of mRNA preheated at 65°C for 5 min served as a template for single-strand cDNA synthesis in a 20 μl reaction mixture containing 3 m M MgCl2, 75 m M KCl, 50 m M Tris-HCl (pH 8.3), 0.5 m M dNTPs, 200 μ M oligo (dT) primer, 20 U of RNase inhibitor, and 200 U of M-MLV reverse transcriptase (Gibco BRL, Gaithersburg, MD, USA) at 37°C for 60 min.
By using multiple templates for a single target, it is possible to generate a model with accuracy higher than any single-template-based models.
Yahoo doesn't have to create the templates for every single search category.
