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PCR: Babe A -D13Mit84 (4.4cM)
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ID:
168
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Amplifluor:
No
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Nucleotide sequence (5´ ⇒ 3´) |
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Primer name |
Sequence |
D13Mit84 F
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TATGTATCCACAGCCCTTAAAATG
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D13Mit84 R
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TGAGTTGCATCCTTGTAACCC
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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:
D13Mit84 F
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6.6 μM
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NA
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1.5
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Primer:
D13Mit84 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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37.45
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Taq Polyermase |
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0.05
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Tritonx100 (10%)
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0.5
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25mM MgCl2
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2.0
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Comments on protocol: |
DNA Sample (ear punch)
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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. |
220
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C57BL/6J
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2. |
242
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NOD
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