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PCR: K14 Flii
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Export PDF
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| ID:
331
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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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Flii F1
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GAAGCCAAGTTGGCAGAAGACATCC
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FLii R1
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TGGAGCGCCTTCTGCAAGGCCCTGGCC
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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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4.5
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| 10x Buffer |
2x ABgene Ready Mix Buffer 4
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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:
Flii F1
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20.0 μM
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NA
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0.25
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| Primer:
FLii R1
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20.0 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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NA
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| Taq Polyermase |
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NA
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| 10x Buffer recipe: |
0.025units/ul Thermoprime Plus DNA polymerase, 1.25ml of 750mM Tris-HCl (pH8.8), 200mM (NH4)2SO4, 0.1% (v/v) Tween 20, 15mM MgCl2, 0.2mM each of dATP, dCATP, dGTP, dTTP, precipitant and red dye. Cat #AB-0AB-8815.
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| Comments on protocol: |
DNA isolated from ear biopsy. Buffer TE-5% Tween, 40ul H2O to 10ul DNA
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| Steps |
Temp (°C ) |
Time (seconds) |
| Hot start:
false
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| Initiation/Melting |
95.0
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180
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| Denaturation |
95.0
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15
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| Annealing |
68.0
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150
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| Elongation |
NA
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Number of cycles
(repeat denature, anneal & elongate):
35
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| Strand completion (i.e. 72°C, 10 min) |
NA
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| Finish (i.e. 4°C, indefinite) |
4.0
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Hold
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| Running buffer |
TBE + SYBR safe
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| % Agarose |
1.5
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| Volt |
80.0
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| Estimated Running Time (min) |
20.0
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| Picture |
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| Number |
Band (bp) |
Genotype |
| 1. |
500
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Transgenic
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