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10.1038/nprot.2014.191 http://scihub22266oqcxt.onion/10.1038/nprot.2014.191 C4327781!4327781 !25675209     free     free     free Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\25675209 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Nat+Protoc 2015 ; 10 (3 ): 442-58 Nephropedia Template TP {{tp|p=25675209 |t=2015. Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression profiling in intact cells and tissues |pdf=|usr=}}{{25675209 }} gab.com Text Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression profiling in intact cells and tissues JO: Nat Protoc http://www.kidney.de/pget.php?&tw=1&pmid=25675209 #FOAvip RNA-sequencing (RNA-seq) measures the quantitative change in gene expression over the whole transcriptome, but it lacks spatial context. In contrast, in situ hybridization provides the location of gene expression, but only for a small number of genes. Here we detail a protocol for genome-wide profiling of gene expression in situ in fixed cells and tissues, in which RNA is converted into cross-linked cDNA amplicons and sequenced manually on a confocal microscope. Unlike traditional RNA-seq, our method enriches for context-specific transcripts over housekeeping and/or structural RNA, and it preserves the tissue architecture for RNA localization studies. Our protocol is written for researchers experienced in cell microscopy with minimal computing skills. Library construction and sequencing can be completed within 14 d, with image analysis requiring an additional 2 d. Twit Text FOAVip Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression profiling in intact cells and tissues ????Nat Protoc http://www.kidney.de/pget.php?&tw=1&pmid=25675209 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25675209 #FOAvip English Wikipedia <ref>{{Cite pmid|25675209 }}</ref> Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression profiling in intact cells and tissues #MMPMID25675209 Lee JH ; Daugharthy ER ; Scheiman J ; Kalhor R ; Ferrante TC ; Terry R ; Turczyk BM ; Yang JL ; Lee HS ; Aach J ; Zhang K ; Church GM Nat Protoc 2015[Mar]; 10 (3 ): 442-58 PMID25675209 show ga RNA-sequencing (RNA-seq) measures the quantitative change in gene expression over the whole transcriptome, but it lacks spatial context. In contrast, in situ hybridization provides the location of gene expression, but only for a small number of genes. Here we detail a protocol for genome-wide profiling of gene expression in situ in fixed cells and tissues, in which RNA is converted into cross-linked cDNA amplicons and sequenced manually on a confocal microscope. Unlike traditional RNA-seq, our method enriches for context-specific transcripts over housekeeping and/or structural RNA, and it preserves the tissue architecture for RNA localization studies. Our protocol is written for researchers experienced in cell microscopy with minimal computing skills. Library construction and sequencing can be completed within 14 d, with image analysis requiring an additional 2 d. |DNA, Complementary/genetics [MESH] |Fluorescence [MESH] |Gene Expression Profiling/*methods [MESH] |Microscopy, Confocal/methods [MESH] |RNA, Messenger/*genetics [MESH]Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression pro(epmc).pdf DeepDyve Pubget Overpricing