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PCR: Vibes B –D15Mit86 (18cM)
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Export PDF
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| ID:
359
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Amplifluor:
No
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| Nucleotide sequence (5´ ⇒ 3´) |
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| Primer name |
Sequence |
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D15Mit86 F
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TACACTTATCTGTTGGAACATCCC
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D15Mit86 R
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ATATAGATAAACACACGCACACCA
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| Reagents / Constituents |
Commercial name |
Stock
Concentration |
20x Primer mix
concentration (μM) |
Volume (μL) |
| DNA Sample |
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NA ng/μl
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1.0
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| 10x Buffer |
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20.0 mM MgCl2
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5.0
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| dNTPs |
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10.0 mM
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1.0
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| 20x amplifluor SNP FAM Primer |
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NA μM
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NA
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| 20x amplifluor SNP JOE Primer |
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NA μM
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NA
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| Primer:
D15Mit86 F
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6.6 μM
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NA
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1.5
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| Primer:
D15Mit86 R
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6.6 μM
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NA
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1.5
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| Amplifluor Primer Volume |
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NA
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| Water |
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38.45
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| Taq Polyermase |
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0.05
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Triton X-100 (10%)
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0.5
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25mM MgCl2
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1.0
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| Comments on protocol: |
DNA Sample (ear punch).
Final concentration of 2.5mM MgCl2
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| Steps |
Temp (°C ) |
Time (seconds) |
| Hot start:
false
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| Initiation/Melting |
94.0
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120
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| Denaturation |
94.0
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20
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| Annealing |
55.0
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20
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| Elongation |
72.0
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20
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Number of cycles
(repeat denature, anneal & elongate):
35
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| Strand completion (i.e. 72°C, 10 min) |
72.0
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180
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| Finish (i.e. 4°C, indefinite) |
10.0
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Hold
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| % Agarose |
3.0
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| Volt |
400.0
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| Estimated Running Time (min) |
20.0
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| Number |
Band (bp) |
Genotype |
| 1. |
172
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C57BL/6
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| 2. |
196
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NOD
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