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PCR: Perforin(prf-/-)
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Export PDF
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| ID:
314
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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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JT272
|
CCACTCCACCTTGACTTCAAAAAGGCG
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JT98
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TGGGCAGCAGTCCTGGTTGGTGACCTT
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01Neo2A
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CGGAGAACCTGCGTGCAATC
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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 |
2x Mix Qiagen multiplex PCR Kit
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NA mM MgCl2
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5.0
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| dNTPs |
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NA mM
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NA
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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:
JT272
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10.0 μM
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NA
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0.25
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| Primer:
JT98
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10.0 μM
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NA
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0.5
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| Primer:
01Neo2A
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10.0 μM
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NA
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0.25
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| Amplifluor Primer Volume |
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NA
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| Water |
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3.0
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| Taq Polyermase |
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NA
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| Comments on protocol: |
DNA Sample (ear punch)
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| Steps |
Temp (°C ) |
Time (seconds) |
| Hot start:
true
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| Initiation/Melting |
95.0
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900
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| Denaturation |
95.0
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20
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| Annealing |
60.0
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30
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| Elongation |
72.0
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60
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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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240
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| Finish (i.e. 4°C, indefinite) |
20.0
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Hold
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| Running buffer |
1.5% sodium borate
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| % Agarose |
2.0
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| Volt |
350.0
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| Estimated Running Time (min) |
20.0
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| Number |
Band (bp) |
Genotype |
| 1. |
975
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Perforin KO/KO
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| 2. |
600
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wt/wt
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