Library |
Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
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Total RNA was separated by 15% agarose gels to extract the small RNA (18-30 nt). After precipitated by ethanol and centrifugal enrichment of small RNA sample, the library was prepared according to the method and process of Small RNA Sample Preparation Kit (Illumina, RS-200-0048). The main contents are as follows: Connecting the 3' adaptor to the separated small RNA; Connecting the 5' adaptor to the separated small RNA;RT-PCR; Recycling strips of 145-160bp (22-30nt RNA). RNA concentration of library was measured using Qubit® RNA Assay Kit in Qubit® 2.0 to preliminary quantify and then dilute to 1ng/μl. Insert size was assessed using the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA), and after the insert size consistent with expectations, qualified insert size was accurate quantitative using Taqman fluorescence probe of AB Step One Plus Real-Time PCR system (Library valid concentration>2nM). The qualified libraries were sequenced by an Illumina platform and 50 bp single-end reads were generated. |
RNA-Seq |
TRANSCRIPTOMIC |
RT-PCR |
SINGLE
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