1 |
CTAB |
2% |
2% |
2.5% |
2.5% |
2 |
Sonication |
3 cycles at high amplitute |
No |
3 cycles at high
amplitute |
No |
3 |
FREEZE-THAW |
3 cycles at 70°C and -20°C |
3 cycles at 70°C and
Liquid Nitrogen |
3 cycles at 70°C and Liquid Nitrogen |
No |
4
|
FREEZE-BOIL
|
No
|
No
|
No
|
3 cycles at 95°C for 5 minutes,
-80°C for 10 minutes
|
5 |
DNase-Free-RNase |
No |
No |
10µl/ml |
10µl/ml |
6 |
Chloroform: isoamyl alcohol (24:1) |
~800 µl |
~800 µl |
~800 µl |
~800
µl |
7 |
Phenol: Chloroform: isoamyl alcohol (25:24:1) |
~600
µl |
~600 µl |
~600 µl |
~600 µl |
8 |
Isopropanol |
~400 µl |
~400 µl |
~400 µl |
~400 µl |
9 |
Ice-cold Ethanol 70% |
1ml, twice |
1ml, twice |
1ml, twice |
1ml,
twice |
10 |
Nucleus Free Water |
30 µl |
30 µl |
30 µl |
30 µl |
11 |
18ssu rRNA PCR |
No |
No |
Yes |
Yes |