Equipment and Materials

Week 1

• NEB Monarch Plasmid Miniprep Kit (New England Biolabs, Ipswich, MA)
• Microcentrifuge at a maximum speed of 13,000 rpm
• NanoDrop™ / One^C Microvolume UV-Vis spectrophotometer (Thermo Scientific, Wilmington, DE)
• Savant™ SpeedVac™ DNA130 Integrated Vacuum Concentrator Systems (Thermo Scientific, Waltham, MA)
• 37°C water bath or incubator
• Ice bucket
• 15 ml Falcon tube
• LB liquid medium
• EcoRI and PstI Restriction enzymes and buffers

Week 2

• Agarose Gel electrophoresis apparatus (Bio-Rad, Hercules, CA)
• Power supply
• 250 ml flask
• 50X Tris-Acetate-EDTA (TAE) buffer for agarose gel electrophoresis
• Agarose (Thermo Fisher Scientific, Waltham, MA)
• GelRed nucleic acid staining dye (Phenix Research Products, Candler, NC)
• LB broth
• Ampicillin
• 1 M NaCl and 0.5 M NaOH solution
• Nylon membrane, Whatman paper, and paper towels
• Containers for wetting, incubating, and treating gels and membranes
• Southern blot assembly
• 5X agarose gel loading buffer
  • 30% glycerol or sucrose (increases sample density)
  • 50 mM Na₂EDTA chelates nucleases
  • 0.25% each of bromophenol blue and xylene cyanol dyes to track the progress of electrophoresis

Week 3

• GeneJET Gel Extraction Kit (Thermo Fisher Scientific, Waltham, MA )
• PCR DIG Probe Synthesis Kit (MilliporeSigma, St. Louis, MO)
• T-100 thermal cycler (Bio-Rad, Hercules, CA)

Week 4

• HYBAID Mini Oven and glass tubes for prehybridization and hybridization (Thermo Electron Corp, Waltham, MA)
• Bench-top shaker
• Water bath set at 60°C
• 2XSSC (saline sodium citrate) 0.1% SDS, and 0.2XSSC 0.1% SDS made from 10% SDS and 20XSSC (3M Sodium chloride, 0.3M Sodium citrate, adjusted pH to 7.0)
• DIG EasyHyb buffer (MilliporeSigma, Burlington, MA)
• DIG Nucleic Acid Detection Kit (MilliporeSigma, Burlington, MA)

Week 5

• Student laptops for web-based sequence analysis