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Status |
Public on Jan 01, 2018 |
Title |
Authentic single cell RNA-Seq by measuring the transcriptome of an individual cell twice |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
We developed a novel single-cell RNA-Seq technique that can directly measure the technical noises for each individual gene by splitting a single cell into two equal halves and measuring their transcriptomes separately (Measure a single cell twice by RNA-Seq, MAST-Seq).The MAST-Seq protocol was developed from the single cell RNA-Seq technique we previously developed (Tang et al. 2010). We first lysated the cell to release the RNA from the cell, then reverse transcripted the mRNA into cDNA, added polyA to the 3' end of cDNA, synthesized the second strand of cDNA used designed primer, and at last PCR amplified the double strand cDNA to get enough amount of DNA to construct the cDNA libraries.
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Overall design |
10 single mouse ES cells, 4 half of single mouse ES cells, 4 quarter of single mouse ES cells, 8 eighth of single mouse ES cells, 4 quarter of 2 single mouse ES cells, 4 quarter of 2 single MEF cells,5 single mouse inner cell mass cells, 5 mouse trophectoderm cells, 4 half of single zygotes, 12 half of single 2-cell embryo blastomeres, 24 half of single 4-cell embryo blastomeres, 8 eighth of 8-cell embryo and 64 half of single 8-cell embryo blastomeres, in total 156 samples.
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Contributor(s) |
Yang L, Ma Z, Cao C, Wu X, Lee R, Hu B, Streets A, Wen L, Huang Y, Lao K, Tang F |
Citation |
Lu Yang, Zhaochun Ma, Chen Cao, Yuhao Zhang, Xinglong Wu, Raymond Lee, Boqiang Hu, Lu Wen, Hao Ge, Yanyi Huang, Kaiqin Lao, and Fuchou Tang. MR-seq: measuring a single cell's transcriptome repeatedly by RNA-seq. Science Bulletin 2017;62(6):391-398. doi:10.1016/j.scib.2017.01.029
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Submission date |
Jan 21, 2015 |
Last update date |
May 15, 2019 |
Contact name |
YangLu Yang |
E-mail(s) |
lucyyang1991@hotmail.com
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Organization name |
Beijing Friendship Hospital, Capital Medical University
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Street address |
Beijing Xicheng District Yongan Road NO.95
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City |
Beijing |
State/province |
Beijing |
ZIP/Postal code |
100053 |
Country |
China |
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Platforms (3) |
GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
GPL18635 |
Ion Torrent Proton (Mus musculus) |
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Samples (156)
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Relations |
BioProject |
PRJNA273643 |
SRA |
SRP052833 |
Supplementary file |
Size |
Download |
File type/resource |
GSE65160_RAW.tar |
101.8 Mb |
(http)(custom) |
TAR (of FPKM_TRACKING) |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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