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A micro-RNA-Seq Method for Transcriptome Analysis of Yak Oocytes
Lan DaoIiang, Xiong Xianrong, Hu...
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Abstract
The RNA -Seq approach has been used as a powerful tool for transcriptome analysis
because it is advantageous over conventional methods. However, this technology requires a large amount of input total RNA. Thus, many research samples such as oocytes are unable to meet the minimum amount request of sequencing. To address this challenge, we established an improved micro-RNA-Seq method in a model of yak oocytes, and the starting amount of total RNA as low as 10 ng. After deep sequencing, a total of 47 619 254 clean reads with 4 Gb data were obtained in the micro-oocyte library. Quality control tests and basic analysis results showed that the sequencing quality of the library was good. Further compared with the standard RNA-Seq method, the result of randomness assessment showed that the reads random distribution in the two sequencing methods looked both well and similar. The correlation coefficient between the QPCR result and micro-RNA-Seq data was high (P = O. 81), and the Pearson correlation of gene expression
level between micro-RNA-Seq and standard RNA-Seq was nearly to O. 9 (P = 0.88), indicating that the micro-RNA-Seq has a high correlation with the standard RNA-Seq. The study provides an improved and reliable transcriptome sequencing method for trace samples, especially for the yak oocyte.
Keywords: Micro-RNA-Seq; oocyte; yak; smart; transcriptome [...]
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About
Affiliation of the authors at the time of publication
Institute of Qinghai-Tibetan Plateau, Southwest University for Nationalities, Chengdu, 610041, China; College of Life Science aod Technology, Southwest University for Nationalities, Chengdu, 610041, China; Beijing Genomics Institute (BGI) -Shenzhen, Shenzhen, 518000, China.
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