| |
|
|
PCR: Stat3
|
Export PDF
|
| ID:
418
|
|
|
Amplifluor:
No
|
|
|
| Nucleotide sequence (5´ ⇒ 3´) |
|
| Primer name |
Sequence |
|
A 1/40
|
AGCAGCTGACAACGCTGGCTGAGAAGCT
|
|
B 1/80
|
TTGCTGCTCTCGCTGAAGCGCAGTAGG
|
|
C 1/80
|
ATCGCCTTCTATCGCCTTCTTGACGAG
|
|
|
|
|
| Reagents / Constituents |
Commercial name |
Stock
Concentration |
20x Primer mix
concentration (μM) |
Volume (μL) |
| DNA Sample |
|
NA ng/μl
|
|
4.0
|
| 10x Buffer |
|
20.0 mM MgCl2
|
|
3.0
|
| dNTPs |
|
10.0 mM
|
|
3.0
|
| 20x amplifluor SNP FAM Primer |
|
NA μM
|
|
NA
|
| 20x amplifluor SNP JOE Primer |
|
NA μM
|
|
NA
|
| Primer:
A 1/40
|
|
10.0 μM
|
NA
|
2.0
|
| Primer:
B 1/80
|
|
10.0 μM
|
NA
|
2.0
|
| Primer:
C 1/80
|
|
10.0 μM
|
NA
|
2.0
|
| Amplifluor Primer Volume |
|
|
|
NA
|
|
| Water |
|
|
|
10.5
|
| Taq Polyermase |
|
|
|
1.0
|
|
25mM MgCl2
|
|
|
|
2.5
|
|
|
|
| Steps |
Temp (°C ) |
Time (seconds) |
| Hot start:
false
|
|
|
| Initiation/Melting |
94.0
|
300
|
| Denaturation |
94.0
|
30
|
| Annealing |
65.0
|
60
|
| Elongation |
72.0
|
60
|
Number of cycles
(repeat denature, anneal & elongate):
40
|
|
|
| Strand completion (i.e. 72°C, 10 min) |
72.0
|
300
|
| Finish (i.e. 4°C, indefinite) |
20.0
|
Hold
|
|
|
|
| % Agarose |
2.0
|
| Volt |
120.0
|
| Estimated Running Time (min) |
60.0
|
| Number |
Band (bp) |
Genotype |
| 1. |
920
|
Mutant
|
| 2. |
1000
|
wt
|
|
