| |
|
|
PCR: B-thy A –D11Mit349 (27.3cM)
|
Export PDF
|
| ID:
163
|
|
|
Amplifluor:
No
|
|
|
| Nucleotide sequence (5´ ⇒ 3´) |
|
| Primer name |
Sequence |
|
D11Mit349 F
|
AGTATCAGAAGATCCAGTTGGAGG
|
|
D11Mit349 R
|
GTAGAAAAAGATACCCAGTGTCAGC
|
|
|
|
|
| Reagents / Constituents |
Commercial name |
Stock
Concentration |
20x Primer mix
concentration (μM) |
Volume (μL) |
| DNA Sample |
|
NA ng/μl
|
|
1.0
|
| 10x Buffer |
|
20.0 mM MgCl2
|
|
5.0
|
| dNTPs |
|
10.0 mM
|
|
1.0
|
| 20x amplifluor SNP FAM Primer |
|
NA μM
|
|
NA
|
| 20x amplifluor SNP JOE Primer |
|
NA μM
|
|
NA
|
| Primer:
D11Mit349 F
|
|
6.6 μM
|
NA
|
1.5
|
| Primer:
D11Mit349 R
|
|
6.6 μM
|
NA
|
1.5
|
| Amplifluor Primer Volume |
|
|
|
NA
|
|
| Water |
|
|
|
39.45
|
| Taq Polyermase |
|
|
|
0.05
|
|
Tritonx100 10%
|
|
|
|
0.5
|
|
| Comments on protocol: |
DNA Sample (ear punch)
|
|
|
| Steps |
Temp (°C ) |
Time (seconds) |
| Hot start:
false
|
|
|
| Initiation/Melting |
94.0
|
120
|
| Denaturation |
94.0
|
20
|
| Annealing |
55.0
|
20
|
| Elongation |
72.0
|
20
|
Number of cycles
(repeat denature, anneal & elongate):
35
|
|
|
| Strand completion (i.e. 72°C, 10 min) |
72.0
|
180
|
| Finish (i.e. 4°C, indefinite) |
10.0
|
Hold
|
|
|
|
| % Agarose |
3.0
|
| Volt |
400.0
|
| Estimated Running Time (min) |
20.0
|
| Number |
Band (bp) |
Genotype |
| 1. |
118
|
C57BL/6J
|
| 2. |
78
|
NOD
|
|
Database
