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10.1101/pdb.prot090803 http://scihub22266oqcxt.onion/10.1101/pdb.prot090803 C4804709!4804709 !26933249     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=26933249 &cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215       Cold+Spring+Harb+Protoc 2016 ; 2016 (3 ): pdb.prot090803 Nephropedia Template TP {{tp|p=26933249 |t=2016. Single Guide RNA Library Design and Construction |pdf=|usr=}}{{26933249 }} gab.com Text Single Guide RNA Library Design and Construction JO: Cold Spring Harb Protoc http://www.kidney.de/pget.php?&tw=1&pmid=26933249 #FOAvip This protocol describes how to generate a single guide RNA (sgRNA) library for use in genetic screens. There are many online tools available for predicting sgRNA sequences with high target specificity and/or cleavage activity. Here, we refer the user to genome-wide sgRNA sequence predictions that we have developed for both the human and mouse and that are available from the Broad Institute website. Once a set of target genes and corresponding sgRNA sequences has been identified, customized oligonucleotide pools can be rapidly synthesized by a number of commercial vendors. Thereafter, as described here, the oligonucleotides can be efficiently cloned into an appropriate lentiviral expression vector backbone. The resulting plasmid pool can then be packaged into lentiviral particles and used to generate knockouts in any cell line of choice. Twit Text FOAVip Single Guide RNA Library Design and Construction ????Cold Spring Harb Protoc http://www.kidney.de/pget.php?&tw=1&pmid=26933249 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26933249 #FOAvip English Wikipedia <ref>{{Cite pmid|26933249 }}</ref> Single Guide RNA Library Design and Construction #MMPMID26933249 Wang T ; Lander ES ; Sabatini DM Cold Spring Harb Protoc 2016[Mar]; 2016 (3 ): pdb.prot090803 PMID26933249 show ga This protocol describes how to generate a single guide RNA (sgRNA) library for use in genetic screens. There are many online tools available for predicting sgRNA sequences with high target specificity and/or cleavage activity. Here, we refer the user to genome-wide sgRNA sequence predictions that we have developed for both the human and mouse and that are available from the Broad Institute website. Once a set of target genes and corresponding sgRNA sequences has been identified, customized oligonucleotide pools can be rapidly synthesized by a number of commercial vendors. Thereafter, as described here, the oligonucleotides can be efficiently cloned into an appropriate lentiviral expression vector backbone. The resulting plasmid pool can then be packaged into lentiviral particles and used to generate knockouts in any cell line of choice. |*CRISPR-Cas Systems [MESH] |*Gene Library [MESH] |Animals [MESH] |Gene Knockout Techniques/*methods [MESH] |Genetic Testing/*methods [MESH] |Humans [MESH] |Mice [MESH]Single Guide RNA Library Design and Construction (epmc).pdf DeepDyve Pubget Overpricing