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PCR: 9B6 35 (amplifluor)
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Export PDF
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ID:
149
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Amplifluor:
Yes
If you require further assistance regarding this PCR protocol, please contact us at genotyping.support@apf.edu.au
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Nucleotide sequence (5´ ⇒ 3´) |
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Primer name |
Sequence |
9B6 35-1
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GAAAGGTCGGAGTCAACGGATTTGTGTAAGCAAACTCGAGCC
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9B6 35-2
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GAAGGTGACCAAGTTCATGCTTCTGTGTAAGCAAACTCGAGCA
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9B6 35-R
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GTGTGTGAGCTGGACTTTGTA
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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 |
10x Reaction Mix S Plus
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NA mM MgCl2
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0.5
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dNTPs |
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2.5 mM
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0.4
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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:
9B6 35-1
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NA μM
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0.5
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Primer:
9B6 35-2
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NA μM
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0.5
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Primer:
9B6 35-R
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NA μM
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7.5
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Amplifluor Primer Volume |
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1.0
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Water |
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1.58
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Taq Polyermase |
Platinum Taq (5U/μl)
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0.05
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Comments on protocol: |
DNA Sample (cleaned ear punch in dH20-see cleanup protocol).
Comments on protocol: Reactions to be done at room temperature with reagents added in order.
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Steps |
Temp (°C ) |
Time (seconds) |
Hot start:
true
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Initiation/Melting |
95.0
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240
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Denaturation |
95.0
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10
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Annealing |
60.0
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20
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Elongation |
72.0
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40
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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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Picture |
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