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PCR: IL13_IL4rMultiplex
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Export PDF
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| ID:
274
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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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IL-13b wt F
|
CAGTCCTGGCTCTTGCTTG
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IL-13b wt R
|
TCTTTGGGGGAACCCTTTAC
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IL-4r wt F
|
CACTCCACACCAATGTGTCC
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IL-4r wt R
|
CCTACCGCTGATTTTTCTCG
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|
neo common F.R
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GGATGATCTGGACGAAGAGC
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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 |
|
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:
IL-13b wt F
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|
10.0 μM
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NA
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0.5
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| Primer:
IL-13b wt R
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|
10.0 μM
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NA
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0.5
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| Primer:
IL-4r wt F
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|
10.0 μM
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NA
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0.2
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| Primer:
IL-4r wt R
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|
10.0 μM
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NA
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0.2
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| Primer:
neo common F.R
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10.0 μM
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NA
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0.5
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| Amplifluor Primer Volume |
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NA
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| Water |
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2.1
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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:
false
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| Initiation/Melting |
95.0
|
900
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| Denaturation |
95.0
|
25
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| Annealing |
58.0
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25
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| Elongation |
72.0
|
90
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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
|
300
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| Finish (i.e. 4°C, indefinite) |
20.0
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Hold
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| % Agarose |
2.0
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| Volt |
360.0
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| Estimated Running Time (min) |
12.0
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| Number |
Band (bp) |
Genotype |
| 1. |
1300
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IL13 KO
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| 2. |
1000
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IL4r KO
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| 3. |
355
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IL4r WT
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| 4. |
299
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IL13 WT
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