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PCR |
| Name: |
Eri_adh1 |
Type: |
Real-Time PCR |
System: |
Quantitative |
| Genetic Element: |
alcohol dehydrogenase 1 |
Ampl. Length: |
81 |
Ampl. Position: |
|
Sequence Size [bp]: |
|
| AnnTemp [°C]: |
60 |
Taxonomy: |
Erica |
Temp. Time Program: |
Process Time [min:s] Temperature [°C]
Pre-PCR: decontamination 02:00 50
Pre-PCR: activation of DNA polymerase and denaturation of template DNA 10:00 95
PCR (45 cycles)
Step 1 Denaturation 00:15 95
Step 2 Annealing and elongation 01:00 60 |
| Device: |
|
Reagents: |
Total volume 10 µl
Template DNA (maximal amount 200 ng) 2 µl
DNA polymerase AmpliTaq Gold® 1,5 IU
Decontamination system dUTP
AmpErase uracil N-glycosylase 400 µmol/l
0,5 IU
Reaction buffer TaqMan® buffer A (containing passive reference ROX)a 1 X
MgCl2 8 mmol/l
Primers 300 nmol/l 300 nmol/l
dNTP dATP, dCTP, dGTP 200 µmol/l each
Probe 200 nmol/l 200 nmol/l
|
nPrimers: |
3 |
Organisation: |
BfR |
| PCR Description: |
|
Sequence: |
|
| LOD [copy]: |
10 |
LOQ [copy]: |
100 |
Efficiency [%]: |
108 |
| Remarks: |
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Primers |
| PPName: |
Type: |
PrimerSequence: |
Length [bp]: |
T_Ann [°C]: |
TmThermo [°C]: |
Tm2AT [°C]: |
GC [%]: |
Position: |
|
Eri_adh1-f |
Sense |
ggA TAg ggg AgT gAT gAT CCA T |
22 |
64.7 (PCR: 60) |
56.0 |
66 |
50.00 |
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Eri_adh1-r |
Antisense |
gAT gTT CCg ACg AAA Tgg TAT ATg |
24 |
65.3 (PCR: 60) |
56.6 |
68 |
41.67 |
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Eri_adh1-probe |
Marker/Probe |
5“- FAM )- TgC CgT TTT TAg AAA ACC TTg ATT TCC CA -(T AMR A)- 3“ |
29 |
72.1 (PCR: 60) |
67.1 |
80 |
37.93 |
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