Journal: Avicenna Journal of Medical Biotechnology

Article Title: Construction and Evaluation of Short Hairpin RNAs for Knockdown of Metadherin mRNA

doi:

Figure Lengend Snippet: Primer sequences for amplification of shDNA cassettes. The underlined nucleotides are HindIII restriction sites

Article Snippet: The expected product was the U6-shDNA fragment that was digested with EcoRI and HindIII restriction enzymes (Jena Bioscience, Germany) and ligated into the pEGFP-V-RS vector (OriGene, USA) by using the T4 DNA ligase enzyme (Thermo Fisher Scientific, USA).

Techniques: Amplification, Sequencing

Journal: Nature protocols

Article Title: Defining genome-wide CRISPR-Cas genome editing nuclease activity with GUIDE-seq

doi: 10.1038/s41596-021-00626-x

Figure Lengend Snippet:

Article Snippet: gRNA preparation RNAse ZAP solution (Fisher Scientific cat. no. AM9780) Plasmid pCRL1 for T7 in vitro transcription of gRNA (Addgene cat. no. 153997) Hind III-HF restriction enzyme (New England Biolabs cat. no. R3104S) HiScribe T7 RNA Synthesis Kit (New England Biolabs cat. no. E2050S) MEGAclear Transcription Clean-Up Kit (Fisher Scientific cat. no. AM1908) RNA High Sensitivity Screen Tape (Agilent cat. no 5067-5579) RNA High Sensitivity Sample Buffer (Agilent cat. no. 5067-5580) Mammalian Cell culture and nucleofection 24 Well Cell Culture Plate, Flat Bottom with Lid, Tissue Culture Treated, Non-Pyrogenic, Polystyrene (Corning Life Sciences cat. no. 3524) Non-Tissue Culture Treated Plate, 24 Well, Flat Bottom with Low Evaporation Lid (Corning Life Sciences cat. no. 351147) Non-Tissue Culture Treated Plate, 96 Well, Flat Bottom with Low Evaporation Lid (Corning Life Sciences cat. no. 351172) Cas9 expressing construct pSQT817 (Addgene cat. no. 53373) gRNA expression construct, backbone BPK1520 (Addgene cat. no. 65777) SE nucleofection kit (Lonza cat. no. V4SC-1096) Advanced DMEM (Fisher Scientific cat. no. 12491023) PBS (Fisher Scientific cat. no. 10010049) Disposable Hemocytometer (Fisher Scientific cat. no. 22600100) Trypan Blue (Fisher Scientific cat. no. 15250061) FBS, Fetal bovine serum (Fisher Scientific cat. no. 10-082-147) Trypsin-EDTA 0.05% (Fisher Scientific cat. no. 25300120) Penicillin/Streptomycin (Fisher Scientific cat. no. 15-070-063) GlutaMax 100X (Fisher Scientific cat. no. 35-050-061) 96 wells plate V-shape (Fisher Scientific cat. no. 720096) X-vivo 15 Culture Media (Lonza BEBP04-744Q) P3 nucleofection kit (Lonza V4SP-3096) !

Techniques: Concentration Assay, Plasmid Preparation

Journal: Nature protocols

Article Title: Defining genome-wide CRISPR-Cas genome editing nuclease activity with GUIDE-seq

doi: 10.1038/s41596-021-00626-x

Figure Lengend Snippet:

Article Snippet: gRNA preparation RNAse ZAP solution (Fisher Scientific cat. no. AM9780) Plasmid pCRL1 for T7 in vitro transcription of gRNA (Addgene cat. no. 153997) Hind III-HF restriction enzyme (New England Biolabs cat. no. R3104S) HiScribe T7 RNA Synthesis Kit (New England Biolabs cat. no. E2050S) MEGAclear Transcription Clean-Up Kit (Fisher Scientific cat. no. AM1908) RNA High Sensitivity Screen Tape (Agilent cat. no 5067-5579) RNA High Sensitivity Sample Buffer (Agilent cat. no. 5067-5580) Mammalian Cell culture and nucleofection 24 Well Cell Culture Plate, Flat Bottom with Lid, Tissue Culture Treated, Non-Pyrogenic, Polystyrene (Corning Life Sciences cat. no. 3524) Non-Tissue Culture Treated Plate, 24 Well, Flat Bottom with Low Evaporation Lid (Corning Life Sciences cat. no. 351147) Non-Tissue Culture Treated Plate, 96 Well, Flat Bottom with Low Evaporation Lid (Corning Life Sciences cat. no. 351172) Cas9 expressing construct pSQT817 (Addgene cat. no. 53373) gRNA expression construct, backbone BPK1520 (Addgene cat. no. 65777) SE nucleofection kit (Lonza cat. no. V4SC-1096) Advanced DMEM (Fisher Scientific cat. no. 12491023) PBS (Fisher Scientific cat. no. 10010049) Disposable Hemocytometer (Fisher Scientific cat. no. 22600100) Trypan Blue (Fisher Scientific cat. no. 15250061) FBS, Fetal bovine serum (Fisher Scientific cat. no. 10-082-147) Trypsin-EDTA 0.05% (Fisher Scientific cat. no. 25300120) Penicillin/Streptomycin (Fisher Scientific cat. no. 15-070-063) GlutaMax 100X (Fisher Scientific cat. no. 35-050-061) 96 wells plate V-shape (Fisher Scientific cat. no. 720096) X-vivo 15 Culture Media (Lonza BEBP04-744Q) P3 nucleofection kit (Lonza V4SP-3096) !

Techniques: Concentration Assay, Plasmid Preparation