Transcriptome data of Litopenaeus vannamei : ovary of female shrimp
1 Total RNA extraction Total RNA was extracted from tissue samples. The concentration and purity of RNA were detected by Nanodrop2000. RNA integrity was detected by agarose gel electrophoresis. RIN value was measured by Agilent2100. A single library requires total RNA  1ug, concentration  35ng / L, OD260 / 280  1.8, OD260 / 230  1.0. 2 Oligo dT Enriched mRNA The 3 ' end of eukaryotic mRNA has the structure of ployA tail. Using magnetic beads with Oligo ( dT ) to perform A-T base pairing with ployA, mRNA can be isolated from total RNA for analysis of transcriptome information. 3 Fragmented mRNA The Illumina Novaseq 6000 platform is used to sequence short sequence fragments. The enriched mRNA is a complete RNA sequence with an average length of several kb, so it needs to be randomly interrupted. With the addition of fragmentation buffer, mRNA can be randomly cleaved and small fragments of about 300 bp can be isolated by magnetic bead screening.4 Reverse synthesis cDNA Under the action of reverse transcriptase, random hexamers were added to reversely synthesize one-stranded cDNA with mRNA as template, and then two-stranded cDNA was synthesized to form a stable double-stranded structure. 5 Connection adaptor The double-stranded cDNA structure is a viscous end, which is complemented by End Repair Mix, and then a ' A ' base is added at the 3 ' end to connect the Y-shaped junction. 6 Sequencing on Illumina platform 1. Library enrichment, PCR amplification for 15 cycles ;  2.2 % agarose gel was used to recover the target band ;  3. TBS380 ( Picogreen ) quantification, mixed in proportion to data ;  4. Bridge PCR amplification was performed on cBot to generate clusters ;  5. Illumina platform sequencing ( PE library, read length 2  150 bp ).