Polymerase Chain Reaction:Article Title: Charting oncogenicity of genes and variants across lineages via multiplexed screens in teratomas
Article Snippet: .. To create the driver ORF library, individual drivers were PCR amplified out of the Cancer Pathways kit (Addgene #1000000072) , individual plasmids (Addgene #9053, #85140, #82262, #82297, #82175, #61852, #39872, #23776, #23688, #10745, #23231) ( ; ; ; ; ; ), a human cDNA pool (Promega Corporation), or obtained as synthesized double-stranded DNA fragments (gBlocks, IDT Inc) with flanking sequences compatible with the BamHI restriction sites. .. The barcoded lentiviral backbone was digested with BamHI HF (New England Biolabs) at 37°C for 3 hours in a reaction consisting of: lentiviral backbone, 4 μg, CutSmart buffer, 5 μl, BamHI, 0.625 μl, H 2 0 up to 50 μl.
Article Title: Charting oncogenicity of genes and variants across lineages via multiplexed screens in teratomas
Article Snippet: .. To create the driver ORF library, individual drivers were PCR amplified out of the Cancer Pathways kit (Addgene #1000000072) , individual plasmids (Addgene #9053, #85140, #82262, #82297, #82175, #61852, #39872, #23776, #23688, #10745, #23231) – , a human cDNA pool (Promega Corporation), or obtained as synthesized double-stranded DNA fragments (gBlocks, IDT Inc) with flanking sequences compatible with the BamHI restriction sites. .. The barcoded lentiviral backbone was digested with BamHI HF (New England Biolabs) at 37 0C for 3 hours in a reaction consisting of: lentiviral backbone, 4 μg, CutSmart buffer, 5 μl, BamHI, 0.625 μl, H 2 0 up to 50 μl.
Amplification:Article Title: Charting oncogenicity of genes and variants across lineages via multiplexed screens in teratomas
Article Snippet: .. To create the driver ORF library, individual drivers were PCR amplified out of the Cancer Pathways kit (Addgene #1000000072) , individual plasmids (Addgene #9053, #85140, #82262, #82297, #82175, #61852, #39872, #23776, #23688, #10745, #23231) ( ; ; ; ; ; ), a human cDNA pool (Promega Corporation), or obtained as synthesized double-stranded DNA fragments (gBlocks, IDT Inc) with flanking sequences compatible with the BamHI restriction sites. .. The barcoded lentiviral backbone was digested with BamHI HF (New England Biolabs) at 37°C for 3 hours in a reaction consisting of: lentiviral backbone, 4 μg, CutSmart buffer, 5 μl, BamHI, 0.625 μl, H 2 0 up to 50 μl.
Article Title: Charting oncogenicity of genes and variants across lineages via multiplexed screens in teratomas
Article Snippet: .. To create the driver ORF library, individual drivers were PCR amplified out of the Cancer Pathways kit (Addgene #1000000072) , individual plasmids (Addgene #9053, #85140, #82262, #82297, #82175, #61852, #39872, #23776, #23688, #10745, #23231) – , a human cDNA pool (Promega Corporation), or obtained as synthesized double-stranded DNA fragments (gBlocks, IDT Inc) with flanking sequences compatible with the BamHI restriction sites. .. The barcoded lentiviral backbone was digested with BamHI HF (New England Biolabs) at 37 0C for 3 hours in a reaction consisting of: lentiviral backbone, 4 μg, CutSmart buffer, 5 μl, BamHI, 0.625 μl, H 2 0 up to 50 μl.
Synthesized:Article Title: Charting oncogenicity of genes and variants across lineages via multiplexed screens in teratomas
Article Snippet: .. To create the driver ORF library, individual drivers were PCR amplified out of the Cancer Pathways kit (Addgene #1000000072) , individual plasmids (Addgene #9053, #85140, #82262, #82297, #82175, #61852, #39872, #23776, #23688, #10745, #23231) ( ; ; ; ; ; ), a human cDNA pool (Promega Corporation), or obtained as synthesized double-stranded DNA fragments (gBlocks, IDT Inc) with flanking sequences compatible with the BamHI restriction sites. .. The barcoded lentiviral backbone was digested with BamHI HF (New England Biolabs) at 37°C for 3 hours in a reaction consisting of: lentiviral backbone, 4 μg, CutSmart buffer, 5 μl, BamHI, 0.625 μl, H 2 0 up to 50 μl.
Article Title: Charting oncogenicity of genes and variants across lineages via multiplexed screens in teratomas
Article Snippet: .. To create the driver ORF library, individual drivers were PCR amplified out of the Cancer Pathways kit (Addgene #1000000072) , individual plasmids (Addgene #9053, #85140, #82262, #82297, #82175, #61852, #39872, #23776, #23688, #10745, #23231) – , a human cDNA pool (Promega Corporation), or obtained as synthesized double-stranded DNA fragments (gBlocks, IDT Inc) with flanking sequences compatible with the BamHI restriction sites. .. The barcoded lentiviral backbone was digested with BamHI HF (New England Biolabs) at 37 0C for 3 hours in a reaction consisting of: lentiviral backbone, 4 μg, CutSmart buffer, 5 μl, BamHI, 0.625 μl, H 2 0 up to 50 μl.
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