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PCR |
| Name: |
Cit_adh1 |
Type: |
Real-Time PCR |
System: |
Quantitative |
| Genetic Element: |
alcohol dehydrogenase 1 |
Ampl. Length: |
83 |
Ampl. Position: |
|
Sequence Size [bp]: |
|
| AnnTemp [°C]: |
60 |
Taxonomy: |
Citrus |
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: |
|
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 300 nmol/l 300 nmol/l<br>dNTP dATP, dCTP, dGTP 200 µmol/l each<br>Probe 200 nmol/l 200 nmol/l<br> |
nPrimers: |
3 |
Organisation: |
BfR |
| PCR Description: |
|
Sequence: |
|
| LOD [copy]: |
0.5 |
LOQ [copy]: |
5 |
Efficiency [%]: |
97 |
| Remarks: |
|
Primers |
| PPName: |
Type: |
PrimerSequence: |
Length [bp]: |
T_Ann [°C]: |
TmThermo [°C]: |
Tm2AT [°C]: |
GC [%]: |
Position: |
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Cit_adh1-f |
Sense |
TgA CAg Agg CgT CAT gCT TAA C |
22 |
67.2 (PCR: 60) |
58.4 |
66 |
50.00 |
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Cit_adh1-r |
Antisense |
TgA ggT TCC AAC gAA ATg ATA AAT A |
25 |
64.7 (PCR: 60) |
56.0 |
66 |
32.00 |
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Cit_adh1-probe |
Marker/Probe |
5Ž -FA M)- CT TgC CAT TgA Tgg AAA ATC TCg ATT TCC -( TAM RA) -3Å ½ |
29 |
72.1 (PCR: 60) |
66.2 |
82 |
41.38 |
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