New nucleic acid molecules and collections thereof, their application and identification
A technology of nucleic acid molecules and aggregates, which is applied in the field of nucleic acid molecules and their aggregates, and can solve problems that have not been fully elucidated
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[0140] Materials and Methods
[0141] Sequencing and analysis of miRNA regions in primates
[0142] Use primer3 software ( http: / / primers.niob.knaw.nl ) designed nested primer sets for PCR amplification of ~700bp regions of 144 known miRNA genes. The choice of primers was based entirely on the human sequence. Genomic DNA from 10 primate species (NIA Senescent Cell Bank DNA Panel PRP00001) (NIA Senescent Cell Bank DNA Panel PRP00001) was purchased from the Coriell Cell Bank (Camden, NJ). All PCR reactions were performed according to the manufacturer's conditions and overall cycle conditions (60 seconds at 94°C, followed by 30 cycles of 94°C for 20 seconds, 58°C for 20 seconds, and 72°C for 60 seconds) in a total volume of 10 μl with 0.5 units of Taq polymerase (Invitrogen, Carlsbad CA). PCR products were sequenced from both ends using an ABI3700 capillary sequencer (Applied Biosystems, Foster City CA). Sequences were quality trimmed and assembled with phred / phrap softwa...
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