Trying Epicentre Illumina kit with midge DNA
Tagmentation
| 14 ul |
water
|
| 1 ul |
1/3 dilution of 162ng/ul DNA extract
|
| 4 μl |
5X Nextera LMW Reaction Buffer
|
| 1 μl |
Nextera Enzyme Mix (Illumina-compatible)
|
- mix on ice
- incubate 5 min @55C
- purify with minelute kit from quiagen and elute into 10 ul buffer
PCR
| 17 μl |
Nuclease-Free water
|
| 5 μl |
Recovered DNA Fragment Library (from Step A3)
|
| 25 μl |
2X Nextera PCR Buffer
|
| 1 μl |
50X Nextera Primer Cocktail
|
| 1 μl |
50X Nextera Adaptor 2*
|
| 1 μl |
Nextera PCR Enzyme (2.5 U/μ
|
- mix on ice
- 72C 3 min; 95C 3min (95C 5s, 62C 30s, 72C 1min)x12, 72C 5 min
- purified using quiagen pcr kit, using 2 washes
- eluted in 50 ul, 6.5ng/ul
Gel
|
Project name
