Reaction setup
|
|
1
Rnx |
5 |
10 |
15 |
20 |
25 |
30 |
35 |
40 |
45 |
50 |
|
H2O |
34.0 |
170 |
340 |
510 |
680 |
850 |
1020 |
1190 |
1360 |
1530 |
1700 |
|
10X
PCR Buffer w/o Mg |
5 |
25 |
50 |
75 |
100 |
125 |
150 |
175 |
200 |
225 |
250 |
|
25
mM MgCl |
5.5 |
27.5 |
55 |
82.5 |
110 |
137.5 |
165 |
192.5 |
220 |
247.5 |
275 |
|
DMSO |
3 |
15 |
30 |
45 |
60 |
75 |
90 |
105 |
120 |
135 |
150 |
|
dNTP,
25mM each |
0.5 |
2.5 |
5 |
7.5 |
10 |
12.5 |
15 |
17.5 |
20 |
22.5 |
25 |
|
tva5,
100pm/ul |
0.25 |
1.25 |
2.5 |
3.75 |
5 |
6.25 |
7.5 |
8.75 |
10 |
11.25 |
12.5 |
|
tva3,
100pm/ul |
0.25 |
1.25 |
2.5 |
3.75 |
5 |
6.25 |
7.5 |
8.75 |
10 |
11.25 |
12.5 |
|
Tag,
5U/UL |
0.3 |
1.50 |
3 |
4.5 |
6 |
7.5 |
9 |
10.5 |
12 |
13.5 |
15 |
|
tail
DNA |
1 |
|
|
|
|
|
|
|
|
|
|
PCR Primers
tva5: CTG CTG CCC GGT AAC GTG ACC GG
tva3: GCC CTG GGG AAG GTC CTG CCC
PCR Program
1. 94C, 5 min
2. 94C, 30 sec
3. 55C, 30 sec
4. 72C, 30 sec
5 30 times to step 2
6. 72C, 10 min
7. 4C for ever.
Run reactions on a 1.5% gel
Product size: 500 bp