Cloning of Small RNAs for the Discovery of Novel MicroRNAs in Plants

Tipo de material: Texto

TextoSeries ; Rice Protocols , 109-118, 2013Trabajos contenidos:

Jagadeeswaran, G
Sunkar, R

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Resumen: Endogenous small RNAs can be grouped into several distinct classes of 21-nt-long microRNAs (miRNAs), short interfering RNAs (siRNAs), trans-acting siRNAs (tasiRNAs), and 24-nt long heterochromatic siRNAs. miRNAs are increasingly being recognized as signi fi cant effectors of gene regulation in a wide range of organisms. These molecules are typically 21-nt long and are amenable for cloning by streamlined protocols. Here we detail the methodology for cloning small RNAs in rice to identify novel miRNAs and other important small RNAs. Brie fl y, small RNA molecules are size fractionated, attached to adaptors, and subsequently converted into cDNA and PCR ampli fi ed. Current high-throughput sequencing technologies allow sequencing of the PCR products directly.

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Previous	B-14693 Specialization and evolution of endogenous small RNA pathways	B-14694 Cross-Talk in Abscisic Acid Signaling	B-14695 Global Identi fi cation of Small RNA Targets in Plants by Sequencing Sliced Ends of Messenger RNAs	B-14696 Cloning of Small RNAs for the Discovery of Novel MicroRNAs in Plants	B-14697 Cloning of Stress-Responsive MicroRNAs and other Small RNAs from Plants	B-14698 CICY: treinta años de labor científica y educativa	B-14699 Other exudates: tragancanth, karaya, mesquite gum and larchwood arabinogalactan	Next

Endogenous small RNAs can be grouped into several distinct classes of 21-nt-long microRNAs (miRNAs), short interfering RNAs (siRNAs), trans-acting siRNAs (tasiRNAs), and 24-nt long heterochromatic siRNAs. miRNAs are increasingly being recognized as signi fi cant effectors of gene regulation in a wide range of organisms. These molecules are typically 21-nt long and are amenable for cloning by streamlined protocols. Here we detail the methodology for cloning small RNAs in rice to identify novel miRNAs and other important small RNAs. Brie fl y, small RNA molecules are size fractionated, attached to adaptors, and subsequently converted into cDNA and PCR ampli fi ed. Current high-throughput sequencing technologies allow sequencing of the PCR products directly.

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