1.1 DNA extraction

1.1 DNA extraction kdorfman Fri, 12/16/2011 - 18:34

Wednesday 1/25/2012

Equipment for 1.1

Equipment for 1.1 kdorfman Mon, 12/19/2011 - 16:27
  • Water bath at 65C
  • Mortar & pestle
  • Dry bath at 65C
  • Centrifuge at 4C
  • 14 mL round bottom POLYPROPYLENE tubes with snap cap. (Becton Dickinson #352059: Fisher 14-959-11B stockroom item)
  • Paper & scotch tape
  • Miracloth (EMD_BIO-475855) & funnels
  • 4-8 pots of densely sown Arabidopsis 1 per pair
  • Medium weighboat for chopped up leaves
  • Paper diaper for bench
  • Liquid N2 + extra ice buckets
  • Stinky waste bucket in hood

Reagents for 1.1

Reagents for 1.1 kdorfman Mon, 12/19/2011 - 17:19

Extracting genomic DNA from Arabidopsis thaliana

For pairs of students
make: aliquots of exactly 6 mL DEB in round-bottom tubes
make: 1 mL aliquots of T10E1
make: 1 mL aliquots of T10E5
make: 1.5 mL aliquots of EtOH 95%
make: 5 mL aliquots of EtOH 70
make: exactly 6 mL aliquots of isopropanol in round bottom tubes
make: 4 mL aliquots of KOAc 5M
make: 100 uL aliquots of NaOAc

Ingredient per pair per class
DEB exactly 6 mL in round-bottomed tube 100 mL
add 5 µL BME per tube at the last minute
T10E1 1 mL 4 mL/pair/semester. Make 100 1mL aliquots
T10E5 1 mL 15 mL
EtOH 95% 1.5 mL 50 mL
EtOH 70% 5 mL 100 mL
ISOprop exactly 6 mL in round-bottom tube 100 mL
KOAc 5M 4 mL 50 mL
NaOAc 3M pH 5.2 100 µL 5 mL