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PCR |
| Name: |
Heli-all-nes |
Type: |
Real-Time PCR |
System: |
Quantitative |
| Genetic Element: |
HAPROFILN allergen |
Ampl. Length: |
106 |
Ampl. Position: |
|
Sequence Size [bp]: |
|
| AnnTemp [°C]: |
56 |
Taxonomy: |
Sunflower: Common Sunflower |
Temp. Time Program: |
Process Time [min:s] Temperature [°C]<br>Pre-PCR: decontamination 02:00 50<br>Pre-PCR: activation of DNA polymerase and denaturation of template DNA 10:00 95<br>PCR (45 cycles) <br>Step 1 Denaturation 00:15 95<br>Step 2 Annealing and elongation 01:00 60 |
| Device: |
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Reagents: |
Total volume 10 µl<br>Template DNA (maximal amount 200 ng) 2 µl<br>DNA polymerase AmpliTaq Gold® 1,5 IU<br>Decontamination system dUTP<br>AmpErase uracil N-glycosylase 400 µmol/l<br>0,5 IU<br>Reaction buffer TaqMan® buffer A (containing passive reference ROX)a 1 X<br> MgCl2 5 mmol/l<br>Primers 900 nmol/l 300 nmol/l<br>dNTP dATP, dCTP, dGTP 200 µmol/l each<br>Probe 200 200 nmol/l<br> |
nPrimers: |
3 |
Organisation: |
BfR |
| PCR Description: |
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Sequence: |
|
| LOD [copy]: |
25 |
LOQ [copy]: |
250 |
Efficiency [%]: |
97 |
| Remarks: |
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Primers |
| PPName: |
Type: |
PrimerSequence: |
Length [bp]: |
T_Ann [°C]: |
TmThermo [°C]: |
Tm2AT [°C]: |
GC [%]: |
Position: |
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Heli-all-nes-f2 |
Sense |
CgT CAA TAC TTg TTA ATA TTA TTA AgA ATT A |
31 |
PCR: 56 |
|
74 |
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Heli-all-r1 |
Antisense |
ATA gCT Tgg CCC gTT TTC TT |
20 |
66.1 (PCR: 56) |
57.3 |
58 |
45.00 |
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Heli-all-nes-probe |
Marker/Probe |
5Ž -FA M)- AT gCA TAT TCC TCC AgC TCC CTg -( TAM RA) -3Å ½ |
23 |
70.0 (PCR: 56) |
61.3 |
70 |
52.17 |
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