small RNA-seq of rice

small RNA-seq of rice Using 500 ng total RNA or enriched small RNA200ng, the 3' splices and 5' splices were connected respectively and then reverse-transcribed using RT primer as primer, using the small RNAs attached to the 5' and 3' splices as templates, cDNA was synthesized under the action of reverse transcriptase. Universal Primer and PCR Primer were used as primers of the two strands respectively, and the first strand of reverse transcriptional cDNA was used as a template to amplify and enrich the fragments. Finally, the glue was cut to recover the small RNA fragments.

水稻小分子RNA测序（small RNA-seq）实验流程为：分别以500 ng总RNA或200 ng富集得到的小分子RNA为起始材料，先将其分别与3'接头、5'接头连接；随后以反转录引物（RT primer）为引物，以带有5'和3'接头的小分子RNA为模板，在反转录酶（reverse transcriptase）的作用下合成互补DNA（cDNA）。之后分别以通用引物（Universal Primer）和PCR引物（PCR Primer）作为两条链的扩增引物，以反转录得到的第一链cDNA为模板，对目标片段进行扩增富集；最终通过切胶回收获取小分子RNA片段。