Week 08 DNA extraction

Dog DNA labs, week of 10/23/23

Isolate DNA

1 sample per pair, from:

• Dog cell cultures from Kathleen
• Student dog mouth swabs
• DNA from Cornell BioBank
• Staff dog mouth swabs

QIAamp DNA minikit 51306

Equipment

• Microfuge tubes
• Sterile forceps (to pull swab from extraction buffer)
• QIAamp columns
• collection tubes
• 56C heat block (10 min)
• vortex mixers
• centrifuge 6000 x g 1 min, 20K x g 3 min
• Nanodrop (may need next class, also) (Need to calculate uL to get 100 ng)

Reagents (per swab)

• PBS (400 UL)
• Proteinase K (20 uL)
• Buffer AL (400 uL)
• 95% EtOH (400 uL)
• QIAamp Mini spin column (1)
• 2 mL collection tube (2?)
• Buffer AW1 (500 uL)
• Buffer AW2 (500 uL)
• Buffer AE (50 uL)

Staff makes spreadsheet on Google Drive with:

• sample # for each dog
• concentration
• quality