Genetics & Evolution lab
Genetics & Evolution lab kdorfman Wed, 10/21/2015 - 22:14Here I have posted the overall proposed DRAFT of the lab schedule for the Genetics and Evolution Lab, as well as write-ups for the individual activities. Any and all critiques/suggestions are welcome. THANKS!
CTAB DNA extraction
CTAB DNA extraction kdorfman Wed, 02/24/2016 - 17:21Materials
Materials kdorfman Wed, 02/24/2016 - 18:28Equipment
- microfuge tubes with blue pestles
- 60C drybath
- ice
- microfuge
Chemicals:
Aliquot 600 µL each:
- CTAB buffer
- isopropanol (ice cold)
Aliquot 750 µL chloroform:isoamyl alcohol.
More than one/student for repeat extraction
CTAB extraction buffer
Recipe
| ingredient | concentration | amount per L |
|---|---|---|
| Tris pH 8 | 1M | 100 mL |
| NaCl | 8.1 % | 81.8 g |
| EDTA | 0.5 M | 40 mL |
| CTAB | 2% | 20 g |
| Beta ME | 0.2% | 2 mL |
Buy it from Fisher: Teknova C2190 CTAB Extraction Solution:
| ingredient | concentration |
|---|---|
| Tris pH 8 | 100 mM |
| NaCl | 1.4 M |
| EDTA pH 8 | 20 mM |
| CTAB | 2% |
Add Beta ME just before aliquoting: 2 µL/mL
Student Protocol
Student Protocol kdorfman Wed, 02/24/2016 - 18:27crush leaf tissue in 600 µL 2X CTAB extraction buffer in microfuge tube using blue pestle
Incubate 60C 30 min. Invert 2x
Add 600 µL chloroform:isoamyl alcohol (24:1)
Centrifuge 5 min
Harvest supernate. (If it's cloudy, repeat chloroform:isoamyl step)
Add equal volume ice cold isopropyl
invert gently until 2 phases mix.
Store at -20C (at least 30 min, better overnight)
