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PCR |
| Name: |
Lav_HMG2 |
Type: |
Real-Time PCR |
System: |
Quantitative |
| Genetic Element: |
Hydroxymethylglutaryl coenzyme A reductase |
Ampl. Length: |
74 |
Ampl. Position: |
|
Sequence Size [bp]: |
|
| AnnTemp [°C]: |
60 |
Taxonomy: |
Lavender (Lavandula lanata) |
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 6 mmol/l<br>Primers 300 300<br>dNTP dATP, dCTP, dGTP 200 µmol/l each<br>Probe 200 200 nmol/l<br> |
nPrimers: |
3 |
Organisation: |
BfR |
| PCR Description: |
|
Sequence: |
|
| LOD [copy]: |
0.5 |
LOQ [copy]: |
5 |
Efficiency [%]: |
117 |
| Remarks: |
|
Primers |
| PPName: |
Type: |
PrimerSequence: |
Length [bp]: |
T_Ann [°C]: |
TmThermo [°C]: |
Tm2AT [°C]: |
GC [%]: |
Position: |
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Lav_HMG2-f |
Sense |
CCA TAT CgC TCg TCT TCA ACA g |
22 |
66.3 (PCR: 60) |
57.6 |
66 |
50.00 |
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Lav_HMG2-r |
Antisense |
TTg CCg gCg ATg gCA |
15 |
69.7 (PCR: 60) |
60.9 |
50 |
66.67 |
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Lav_HMG2-f-probe |
Marker/Probe |
5Ž -FA M)- CA CTT gAT gCT CTg AAg CTT ggC gA -(T AMR A)- 3Ž |
25 |
72.1 (PCR: 60) |
66.2 |
76 |
52.00 |
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