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Always with updated information - www.nuncbrand.comNucleolink Plates, Modules and StripsProteomics & Genomics• Covalent heat-stable binding of DNAAvailable in three formats:--C96 MicroWell Plate--Break Apart Modules--1 x 8 well StripsNucleoLink Plates and Modules:• Optimized for DNA hybridization assays• Available in transparent only• Break Apart format provides maximumflexibility• All modules available with color codingon requestNucleoLink Strips:The DIAPOPSTechniqueBPrimer OnePrimer TwoTemplateDetectionProbeDetection of Immobilized AmplifiedProducts using NucleoLink Strips ina One Phase System• Available in three colors:--Black for fluorescence assays--White for luminescence assays--Transparent for colorimetric assays• Optimized for Solid Phase PCR anddetection in the same well (DIAPOPS)--Ideal for hybridization assays1) BindingPrimer “One” isbound covalentlyto the surface of awell. For reasons ofsimplicity we havenamed the primers“One” and “Two”. Inthe actual assay the bound oligonucleotide canbe either the “upstream” or the “downstream”primer of an amplifica tion sequence.2) Additions ofReagentsBuffer, nucleo tides,Taq poly merase,Template, primer“One” and primer“Two” are added tothe liquid phase. Inthe liquid phase the ratio between primer “One”and primer “Two” should be 1:8.Features:• Sealable with heat-stable Tape 8• Thin walls (0.35 mm)• V-shaped wells with readable flatbottoms• Compatible with standard equipmentin MicroWell plate format includingmost 0.2 ml thermal cycler formats• Resists temperatures up to +121°C3) AmplificationAmplification is initiatedin the liquidphase. The ampliconsin the liquid phasewill hybri di ze withthe bound primermole cules. Theseprimer molecules will be extended by the Taqpolymerase.5) DenaturationAmplicons in theliquid phase areremoved by washing.The bound ampliconsare converted to single-strandedmolecules(denatured)by treatment with NaOH.4) Two types ofAmpliconAfter amplificationthe well contains twotypes of amplicon:Those in the liquidphase and thosebound to the well.6) DetectionThe single-strandedmolecules can bedetected by hybridizationwith a detectionprobe. When“Elisa-like” proceduresare used fordetection, the result can be read in a colorimetricor fluorometric 96 well plate reader.BDIAPOPS:• “ELISA-like” procedure combining solidphase PCR and detection by hybridizationin the same well• Replace conventional and timeconsuming detection methods suchas gel electrophoresis and Southernblotting• No transfer from amplification systemto detection system is necessary,reducing the risk of contaminationLiterature:Tech Note Nos: 17, 18, 19 Vol. 5., 36, 37See Pages 195-196 for full reference list with titles and links106