Nurturing Life Sciences
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#Id: 2611
#Unit 13. Methods in Biology
RACE can be used to amplify unknown 5' (5'-RACE) or 3' (3'-RACE) parts of RNA molecules where part of the RNA sequence is known and targeted by a GSP gene-specific primer.
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Our on-going research indicates that RLMRACE, PPM-RACE, and qRT-PCR are very effective in the verification of sequences of miRNA targets obtained by Degradome sequencing.
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The protocol for RLM-RACE, PPMRACE, and qRT-PCR is rapid, effective, cheap, and can be completed within 2–3 day.
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To more accurately validate the predicted target genes of miRNAs and exactly determine the cleavage sites within the targets, an integrated strategy comprising RLM-RACE, Poly(A) Polymerase-Mediated (PPM)-RACE, and qRT-PCR was developed.
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In plants, the perfect complementarity between most miRNAs and their targets enables the accurate predictions of their targets, while slicing of the targeted mRNAs facilitates target validation through RNA Ligase-Mediated (RLM)-Rapid Amplification of cDNA Ends (RACE) method.
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Type I restriction endonucleases are complex endonucleases and have recognition sequences of about 15 bp;