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PCR

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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: Reagents: Total volume 10 µl<br>Template DNA (maximal amount 200 ng) 2 µl<br>DNA polymerase AmpliTaq Gold&reg; 1,5 IU<br>Decontamination system dUTP<br>AmpErase uracil N-glycosylase 400 µmol/l<br>0,5 IU<br>Reaction buffer TaqMan&reg; 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: Sequence:
LOD [copy]: 25 LOQ [copy]: 250 Efficiency [%]: 97
Remarks:

Food/Product

Food Name: Generic Image: Product Type: Food Source: Product Website:
Miel de CorseSUGAR AND HONEY [A0462]INSECT [B1220]http://europa.e... /en/1129_en.htm

Primers

PPName: Type: PrimerSequence: Length [bp]: T_Ann [°C]: TmThermo [°C]: Tm2AT [°C]: GC [%]: Position:
Heli-all-nes-f2 Sense CgT CAA TAC TTg TTA ATA TTA TTA AgA ATT A 31 PCR: 56 74
Heli-all-r1 Antisense ATA gCT Tgg CCC gTT TTC TT 20 66.1 (PCR: 56) 57.3 58 45.00
Heli-all-nes-probe Marker/Probe 5Ž -FA M)- AT gCA TAT TCC TCC AgC TCC CTg -( TAM RA) -3 23 70.0 (PCR: 56) 61.3 70 52.17

Publications (more...)

Short Title Type Date Download
SOP PlantSpecies by PCR SOP September 2007 SOP_BfR_species-detection_071016.pdf
Analytical Methods: Detection of plant species in honey Journal Accepted: 22.09.2008 FoodChemistry_118_979-986_Laube_Honey_2010.pdf

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