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PCR: To Distinguish the Cited2 wildtype allele from C2F Conditional Allele
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Export PDF
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| ID:
135
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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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C2fF
|
GTCTCAGCGTCTGCTCGTTT
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C2fR
|
CTGCTGCTGTTGGTGATGAT
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| Reagents / Constituents |
Commercial name |
Stock
Concentration |
20x Primer mix
concentration (μM) |
Volume (μL) |
| DNA Sample |
|
NA ng/μl
|
|
1.0
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| 10x Buffer |
|
15.0 mM MgCl2
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|
2.0
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| dNTPs |
|
2.5 mM
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|
2.0
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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:
C2fF
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|
20.0 μM
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NA
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1.0
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| Primer:
C2fR
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|
20.0 μM
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NA
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1.0
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| Amplifluor Primer Volume |
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NA
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| Water |
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11.5
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| Taq Polyermase |
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0.5
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| Steps |
Temp (°C ) |
Time (seconds) |
| Hot start:
false
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| Initiation/Melting |
95.0
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60
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| Denaturation |
95.0
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30
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| Annealing |
58.0
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30
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| Elongation |
72.0
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30
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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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600
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| Finish (i.e. 4°C, indefinite) |
11.0
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| % Agarose |
2.0
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| Volt |
80.0
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| Estimated Running Time (min) |
40.0
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| Number |
Band (bp) |
Genotype |
| 1. |
210
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C2F
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| 2. |
166
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C2 WT
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