HotSHOT genomic DNA prep

For fin clip, tail snip, or ear punch

• Obtain tissue 2 mm
  • tail snip
  • ear punch
  • fin clip
• Place tissue in 96 well plate
• Add 75 uL Alkaline Lysis Reagent for mouse, 50 uL for fish
• Heat to 95C 30 min (10 min to 1 hr)
• Cool to 4C
• Add 75 uL Neutralization Buffer for mouse, 50 uL for fish
• Use 3 uL (1-5) pre PCR reaction

Notes:

• DNA is suitable for PCR, not Southerns
• Heating longer than 30 min does not increase [DNA]
• pH of reagents does not need to be altered
• DNA yield is similar for tail snips and ear punches
• Too much tissue destroys PCR
• Store DNA at 4C or 20C