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SRX1065256: GSM1715563: in_vivo_group (conventionally bred - RNA); Sus scrofa; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 15M spots, 732.6M bases, 406.1Mb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Genome-wide gene expression and DNA methylation reveals Genetic and Epigenetic differences in cloned piglets and conventionally bred normal piglets (RNA-Seq)
show Abstracthide Abstract
Somatic cell nuclear transfer has brought considerable chances to breed excellent breeds and protect endanger animals, while also produced numerous fail embryos and abnormal individuals due to inefficient epigenetic modification at the same time. To understand some mechanisms of abnormal piglets with phenotypes such as macroglossia, standing and walking disabilities in our study and find some differences between abnormal piglets and conventionally bred normal piglets, DNA methylation profile and genome-wide gene expression were conducted in two groups, using methylated DNA immunoprecipitation binding highthroughput sequencing (MeDIP-Seq) and RNA sequencing(RNA-Seq). We generated and provided a genome-wide DNA methylation and gene expression profile for abnormal cloned and conventionally bred piglets. We detected a total of 1493 genes differentially expressed in two groups and 382 of these genes also differentially methylated in two groups. Analysis of relationship between DNA methylation and gene expression revealed that DNA methylation levels had significantly negative and monotonic correlation with gene expression levels in particular regions of genes while no obvious monotonic correlation in other regions. Besides, we found some interesting genes and pathways such as MYH7 and mTOR signalling pathway that may played essential role in muscle growth and development. Briefly, these results provide reliable data for future epigenetic studies and may help to uncover the mechanism of failure clones via SCNT. Overall design: We dissected the leg muscle from the cloned piglets and the conventionally bred piglets, and analyzed the difference of MeDIP-seq and RNA-seq between the two groups. As for data of abnormal cloned piglets, we downloaded it from GEO under Super-Series accession No. GSE51477, including SubSeries accession No.GSE51282 for RNA-seq data (No. GSM1241829 for abnormal cloned group) and SubSeries accession No. GSE51476 for MeDIP-seq data (No. GSM1246252 for abnormal cloned group).
Sample: in_vivo_group (conventionally bred - RNA)
SAMN03782154 • SRS965151 • All experiments • All runs
Organism: Sus scrofa
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was extracted from samples of biceps femoris muscle using TRIzol Reagent (Invitrogen). The concentration and quality of RNA was confirmed using an Agilent 2100 system and agarose gel electrophoresis (AGE). Equal amounts of RNA from the tissue samples of three abnormal cloned piglets, three normal cloned piglets were mixed into three pools. The two independent RNA pools were then used for library construction and sequencing by the Beijing Genome Institute. Briefly, after evaluation of the quality of total RNA, mRNA was enriched by using oligo(dT) magnetic beads. After adding the fragmentation buffer, mRNA was interrupted to short fragments (˜200 bp). First-strand cDNA was synthesized using a random hexamer primer using the mRNA fragments as templates. Then buffer, dNTPs, RNase H and DNA polymerase I were added to synthesize the second strand. Double-strand cDNA was purified with a QiaQuick PCR Extraction kit and washed with Ethidium Bromide buffer for end repair and addition of single nucleotide A (adenine). Finally, sequencing adaptors were ligated to the fragments. Required fragments were purified by AGE and enriched by PCR amplification.
Links:
Runs: 1 run, 15M spots, 732.6M bases, 406.1Mb
Run# of Spots# of BasesSizePublished
SRR206994614,951,003732.6M406.1Mb2016-02-26

ID:
1544554

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