Journal: Journal of visualized experiments : JoVE

Article Title: Analysis of Somatic Hypermutation in the JH4 Intron of Germinal Center B cells from Mouse Peyer’s Patches

doi: 10.3791/61551

Figure Lengend Snippet: Materials

Article Snippet: ​ Reagent Catalog # Vendor Gibco phosphate buffered saline PBS pH 7.4 10-010-049 Fisher Fetal bovine serum S11150 R&D Systems (Atlanta Biologicals) Tissue culture plate 6 well, sterile 08-772-1B Fisher Falcon cell strainer 40μm 08-771-1 Fisher BD slip tip 1mL syringe, sterile 14-823-434 Fisher Corning Falcon test tube with cell strainer snap cap 08-771-23 Fisher Falcon round-bottom polystyrene pubes (FACS tubes) 14-959-5 Fisher Unlabeled anti-mouse CD16/CD32 (Fc block), Clone 2.4G2, BD BDB553142 Fisher Biotinylated peanut agglutinin (PNA) B-1075-5 Vector Labs Streptavidin APC-eFluor 780 Conjugate, eBioscience 47-4317-82 eBioscience DAPI (4′,6-Diamidino-2-Phenylindole, dihydrochloride) (0.5mg/ml) D1306 Fisher PE anti-CD45R/B220, clone RA3-6B2 553090 BD APC-eFluor780 anti-CD45R/B220, clone RA3-6B2, eBioscience 47-0452-80 eBioscience QIAquick Gel Extraction Kit 28706 Qiagen Q5 High-Fidelity DNA Polymerase M0491L NEB 0.2 ml PCR 8-tube FLEX-FREE strip, attached clear flat caps, mixed 1402-4708 USA Scientific Thermo Scientific CloneJET PCR Cloning Kit FERK1231 ThermoFisher ElectroMAX DH10B competent cells 18-290-015 Fisher dNTP N0447L NEB Ampicillin sodium salt BP1760-5 Fisher Seal-Rite 1.5mL microcentrifuge tubes 1615-5500 USA Scientific Falcon™ round-bottom polystyrene tubes (capped) 149591A Fisher Proteinase K BP1700-100 Fisher Glycogen 10901393001 Sigma T4 DNA ligase M020L NEB C57BL/6J mice 000664 Jackson Laboratories Open in a separate window Materials

Techniques: Saline, Sterility, Blocking Assay, Plasmid Preparation, Gel Extraction, Stripping Membranes, PCR Cloning