Abstrakt
Eines der wichtigsten Probleme in Bezug auf die Leistung von PCR-Verbrauchsmaterialien (Platten, Streifen und entsprechende Versiegelungsoptionen) ist ihre Kompatibilität mit mehreren auf dem Markt erhältlichen Cycler-Systemen. In dieser Application Note wurden Eppendorf PCR/qPCR Consumables auf Kompatibilität mit den wichtigsten Cycler-Marken durch einen Physical-Fit-Test mit den Cycler-Blöcken sowie durch exemplarische PCR- und qPCR-Assays für einige Instrumente getestet.
Einführung
PCR- und qPCR-Verbrauchsmaterialien unterscheiden sich in zahlreichen Parametern. Darunter spielen Materialeigenschaften und -qualität, Brunnen-/Schürzenabmessungen, Brunnendicke und Oberflächenbeschaffenheit die wichtigste Rolle. Diese Unterschiede führen zusammen mit verschiedenen Versiegelungsoptionen zu oft erheblichen Abweichungen beim Wechsel von einer Verbrauchsartikelmarke zu einer anderen und können die Gesamtleistung des PCR/qPCR-Assays beeinflussen. Daher ist es von großer Bedeutung festzustellen, ob ein bestimmtes Verbrauchsmaterial mit einer bestimmten PCR/qPCR-Plattform kompatibel ist. Hier präsentieren wir Ihnen einen Überblick über die Eppendorf twin.tec PCR Plates, Eppendorf PCR Tube Strips sowie Eppendorf Cap Strips und ihre Kompatibilität mit den wichtigsten derzeit auf dem Markt befindlichen Cycler-Marken.
Materialen und Methoden
Kompatibilität mit Standard-PCR- und qPCR-Geräten In den meisten Fällen wurde die Kompatibilität der Eppendorf PCR Consumables durch ihre Passung in den entsprechenden Cycler-Block in Kombination mit dem Verschluss des Cycler-Deckels bestimmt. Einige Systeme wurden auch auf ihre Leistungsfähigkeit in einem PCR- oder qPCR-Assay getestet.
Natürlich sind diese Experimente nicht repräsentativ. Daher wird es für jeden Anwender immer notwendig sein, die Verbrauchsmaterialien in seiner eigenen Anwendung zu testen. PCR-Assay: Amplifikation von gDNA (Promega) in einer PCR. Nach dem PCR-Assay wurden die Proben mittels Agarose-Gelelektrophorese (E-Gel, Invitrogen) aufgetrennt und dokumentiert.
Kompatibilität und Testleistung wurden anhand des Vorhandenseins und der Qualität von PCR-Produkten auf dem Gel bewertet (Daten nicht gezeigt).
for BioRad i-Serie, Stratagene Mx, ABI 3000 (excl. adapter) |
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A28242L | GeneOn | 5x25 pcs | 363.6 EUR |
ABI 3000, 7000; BioRad i-Serie, Stratagene Mx, Eppendorf Mastercycler |
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A28239L | GeneOn | 5x25 pcs | 372 EUR |
Stratagene Mx (incl. Perfect Fit Frames from Stratagene), Biorad CFX 96 |
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A28240L | GeneOn | 5x25 pcs | 346.8 EUR |
Sequencer; ABI 3000, 7000 7900HT; Stratagene Mx4000 and other |
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A20232L | GeneOn | 5x25 pcs | 372 EUR |
Universal Stratus Adapter |
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MIC6036 | Scientific Laboratory Supplies | EACH | 592.8 EUR |
BioRad CFX96, Megabase 500/1000 |
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A28241L | GeneOn | 5x25 pcs | 363.6 EUR |
Universal adapter for 5mL tube (58 series) set of 8 |
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E5820732008 | Scientific Laboratory Supplies | EACH | 27.6 EUR |
Adapter for 8x5 mL tubes for rotor S-4-72 (58 series) set of 2 |
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E5804793005 | Scientific Laboratory Supplies | EACH | 207.6 EUR |
Adapter for 1x5 mL tubes for rotor F-34-6-38 (58 series) set of 2 |
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E5804777000 | Scientific Laboratory Supplies | EACH | 90 EUR |
Adapter for 1x5 mL tubesfor rotor FA-45-6-30 (58 series) setof 2 |
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E5820730005 | Scientific Laboratory Supplies | EACH | 90 EUR |
Adapter for 1x5.5-2mL tubes for rotor (58 series) FA-45-20-17 set of 4 |
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E5820768002 | Scientific Laboratory Supplies | EACH | 37.2 EUR |
Adapter for cryogenic tubes for rotor FA-45-20-17 (58 series) setof 4 |
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E5820769009 | Scientific Laboratory Supplies | EACH | 36.72 EUR |
Adapter for auto sampler vials for rotor FA-45-20-17 (58 series) set of 4 |
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E5820770007 | Scientific Laboratory Supplies | EACH | 39.6 EUR |
Adapter for Safe-Lock Tube 5.0 mL for rotor A-4-38 (5702 series) round buckets set of 2 |
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E5702733008 | Scientific Laboratory Supplies | EACH | 97.2 EUR |
Adapter Xk50 |
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28989880 | Scientific Laboratory Supplies | EACH | 525.6 EUR |
Flow Adapter |
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GAS1324 | Scientific Laboratory Supplies | EACH | 102 EUR |
QUICKFIT(R) STRAIGHT TUBING ADAPTER JOI |
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Z322237-5X1EA | Scientific Laboratory Supplies | PK5 | 171.6 EUR |
Adapter Syringe |
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DEN1010 | Scientific Laboratory Supplies | EACH | 56.4 EUR |
Stacking Adapter |
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480158 | Scientific Laboratory Supplies | EACH | 226.8 EUR |
Biosart 100 Monitor Adapter |
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16424 | Scientific Laboratory Supplies | EACH | 38.4 EUR |
Extension Adapter |
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EVA5034 | Scientific Laboratory Supplies | EACH | 52.8 EUR |
Ac Adapter UK |
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F30705 | Scientific Laboratory Supplies | EACH | 82.8 EUR |
Adapter 80 - 120mL 2pcs |
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E5825723007 | Scientific Laboratory Supplies | PK2 | 207.6 EUR |
0.2ml Adapter pack |
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C1008-A2-9 | Benchmark Scientific | 1 PC | 475.15 EUR |
DIN plug adapter |
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PHM1251 | Scientific Laboratory Supplies | EACH | 330.32 EUR |
BNC plug adapter |
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PHM1252 | Scientific Laboratory Supplies | EACH | 330.32 EUR |
Adapter 17.5 x 75-100mm |
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E5920708007 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
Tank level adapter |
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ZSTWIN0A1 | Scientific Laboratory Supplies | EACH | 181.48 EUR |
Adapter for 17.5x100mm |
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E5910709005 | Scientific Laboratory Supplies | PK2 | 267.6 EUR |
Adapter for HOM2518 |
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HOM2520 | Scientific Laboratory Supplies | EACH | 267.6 EUR |
Universal AC adapter |
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PIP3214 | Scientific Laboratory Supplies | EACH | 78 EUR |
MM-Separator PCR strip adapter |
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MD90003 | AMSBIO | 1 | 192 EUR |
Adapter 17.5 x 100mmhighcapacity |
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E5920722000 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
PD-10 Spin Adapter |
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60506-5Pieces | AAT Bioquest | 5 Pieces | 79 EUR |
PD-10 Spin Adapter |
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60507-10Pieces | AAT Bioquest | 10 Pieces | 99 EUR |
Adapter D12.6mm 1.0ml Cryo |
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CEN2624 | Scientific Laboratory Supplies | PK6 | 156 EUR |
Adapter D12.6mm 1.8ml Cryo |
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CEN2626 | Scientific Laboratory Supplies | PK6 | 156 EUR |
Adapter Pediatric-tubes |
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CEN0173 | Scientific Laboratory Supplies | PK6 | 130.8 EUR |
Adapter 50mL highcapacity |
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E5920715003 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
Replacement power adapter |
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HOM2748 | Scientific Laboratory Supplies | EACH | 378.62 EUR |
Adapter 16 x 75-100mm highcap |
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E5920720007 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
Adapter 750mL bottle rotor set |
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E5910757000 | Scientific Laboratory Supplies | PK2 | 296.4 EUR |
Adapter 500mL bottle rotor set |
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E5910760000 | Scientific Laboratory Supplies | PK2 | 296.4 EUR |
Adapter D8mm 0.5ml Tubes |
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CEN2618 | Scientific Laboratory Supplies | PK6 | 50.4 EUR |
Adapter D13.5mm 7ml Tubes |
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CEN2636 | Scientific Laboratory Supplies | PK2 | 61.2 EUR |
Adapter D13.5mm 5ml Tubes |
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CEN2638 | Scientific Laboratory Supplies | PK2 | 52.8 EUR |
Adapter 48x1.5/2.0ml D11mm |
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CEN2666 | Scientific Laboratory Supplies | PK2 | 488.4 EUR |
Adapter 8x1.5mL D11mm 2PK |
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CEN2760 | Scientific Laboratory Supplies | PK2 | 224.4 EUR |
Adapter for 50mL tubes |
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E5910712006 | Scientific Laboratory Supplies | PK2 | 248.4 EUR |
MM-Separator PCR strip adapter P |
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MDMG0003 | AMSBIO | 1 | 192 EUR |
Adapter 1x50ml D30mm FA |
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CEN2660 | Scientific Laboratory Supplies | PK2 | 224.4 EUR |
Adapter 4x15ml D17mm FA |
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CEN2662 | Scientific Laboratory Supplies | PK2 | 224.4 EUR |
Adapter 1x15ml D17mm RB |
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CEN2670 | Scientific Laboratory Supplies | PK2 | 87.6 EUR |
Adapter for 35mm tissue |
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3510148 | Atto | 1unit | 6697.2 EUR |
Adapter D26mm 30ml Tubes |
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CEN2640 | Scientific Laboratory Supplies | PK2 | 124.8 EUR |
Adapter D18mm 16ml Tubes |
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CEN2642 | Scientific Laboratory Supplies | PK2 | 124.8 EUR |
Adapter D17mm 15ml Tubes |
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CEN2644 | Scientific Laboratory Supplies | PK2 | 124.8 EUR |
InSitu-1 Plate Adapter |
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InSitu-01CL-PA1 | MiTeGen | - | 62 EUR |
Adapter 1.6 - 5 ml tubes |
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CEN0189 | Scientific Laboratory Supplies | PK6 | 136.8 EUR |
0.2ml Adapter pack, 6/pk. |
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C1008-A2-6 | Benchmark Scientific | 1 PC | 26.2 EUR |
0.2ml Adapter pack, 8/pk. |
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C1008-A2-8 | Benchmark Scientific | 1 each | 33 EUR |
0.5ml Adapter pack, 6/pk. |
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C1008-A5-6 | Benchmark Scientific | 1 PC | 26.2 EUR |
0.5ml Adapter pack, 8/pk. |
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C1008-A5-8 | Benchmark Scientific | 1 PC | 33 EUR |
12v vehicle power adapter |
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BLO1258 | Scientific Laboratory Supplies | EACH | 81.6 EUR |
12v vehicle power adapter |
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BSH100-A12 | Benchmark Scientific | 1 PC | 58.7 EUR |
Adapter for 11x75-100mm |
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E5910704003 | Scientific Laboratory Supplies | PK2 | 248.4 EUR |
Adapter for 12x75mm tubes |
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E5910711000 | Scientific Laboratory Supplies | EACH | 267.6 EUR |
Adapter for 17.5x100mm tubes |
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E5910762003 | Scientific Laboratory Supplies | PK2 | 319.2 EUR |
Adapter 15 mL highcapacity |
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E5920716000 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
Rubber adapter sleeve 15ml |
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TUB2720 | Scientific Laboratory Supplies | PK2 | 79.2 EUR |
Adapter 13 x 75-100mm high-cap |
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E5920718002 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
Adapter for 250 mL bottle |
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E5910705000 | Scientific Laboratory Supplies | PK2 | 267.6 EUR |
Adapter for 400 mL bottle |
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E5910706006 | Scientific Laboratory Supplies | PK2 | 170.4 EUR |
Transcriptional adapter 3 Antibody (Biotin) |
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20-abx105989 | Abbexa |
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Adapter 250 mL bottles high-cap |
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E5920717006 | Scientific Laboratory Supplies | EACH | 319.2 EUR |
Adapter Molecule Crk (CRK) Antibody (Biotin) |
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20-abx334580 | Abbexa |
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Kinesis Injection Port Adapter; 10-32 |
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CHR3629 | Scientific Laboratory Supplies | EACH | 55.2 EUR |
Adapter D6mm 0.2/0.4ml Tubes |
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CEN2620 | Scientific Laboratory Supplies | PK6 | 50.4 EUR |
Adapter for 24 well plate |
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3510146 | Atto | 1unit | 6697.2 EUR |
Adapter for 96 well plate |
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3510147 | Atto | 1unit | 6697.2 EUR |
Adapter for 4-15ml Tubes |
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GBA1763 | Scientific Laboratory Supplies | EACH | 180 EUR |
Adapter for 5/15mL tubes |
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E5910702000 | Scientific Laboratory Supplies | PK2 | 267.6 EUR |
Adapter Hemogard-cap tubes |
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CEN0175 | Scientific Laboratory Supplies | EACH | 136.8 EUR |
Adapter tubes/ bottles/ plates |
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E5910751001 | Scientific Laboratory Supplies | PK2 | 448.8 EUR |
Screwthread Adapter B14 Cone |
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QMF10/1B/SC | Scientific Laboratory Supplies | EACH | 28.8 EUR |
STE20-Related Kinase Adapter Protein Alpha (STRADA) Antibody |
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20-abx013673 | Abbexa |
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CUBE Cassette Carrier Adapter |
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LFRCA-001 | BioAssayWorks | Each | 631.2 EUR |
Goniometer Head Adapter (Large) |
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GHA-2 | MiTeGen | 1 UNIT | 62 EUR |
Grant-Bio Bluetooth Adapter |
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SHA7808 | Scientific Laboratory Supplies | EACH | 58.8 EUR |
Adapter D11mm 1.5/2.0ml Tubes |
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CEN2622 | Scientific Laboratory Supplies | PK6 | 99.6 EUR |
Adapter 1x50mL D29.5mm FA 2PK |
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CEN2750 | Scientific Laboratory Supplies | PK2 | 195.6 EUR |
Adapter 4x30mL D25.5mm RB 2PK |
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CEN2754 | Scientific Laboratory Supplies | PK2 | 195.6 EUR |
Tube Adapter 10mm and 12mm |
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COL3418 | Scientific Laboratory Supplies | EACH | 63.6 EUR |
Adapter 3x50mL D29mm RB 2PK |
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CEN2752 | Scientific Laboratory Supplies | PK2 | 195.6 EUR |
Adapter 6x15mL D17mm FA 2PK |
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CEN2756 | Scientific Laboratory Supplies | PK2 | 195.6 EUR |
Adapter 5x10mL D16mm RB 2PK |
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CEN2762 | Scientific Laboratory Supplies | PK2 | 224.4 EUR |
Adapter 2x30mL D26mm RB 2PK |
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CEN2764 | Scientific Laboratory Supplies | PK2 | 224.4 EUR |
Adapter 1x50mL D29mm RB 2PK |
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CEN2766 | Scientific Laboratory Supplies | PK2 | 224.4 EUR |
IKA ROTAVISC DIN Adapter Set |
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VIS2144 | Scientific Laboratory Supplies | EACH | 2355.6 EUR |
Adapter for 1.6 - 7 ml tubes |
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A1783 | Scientific Laboratory Supplies | EACH | 180 EUR |
qPCR-Assay: Für die Leistung des qPCR-Assays wurde ein Standardassay mit Lambda-DNA (Promega) und KAPA SYBR Fast qPCR Mastermix (KAPA Biosystems) verwendet. Drei unabhängige qPCR-Assays über sechs Konzentrationsprotokolle der Lambda-DNA-Matrize (101-106) und 3 Wiederholungen für jede Konzentration wurden durchgeführt. Die Kompatibilität wurde basierend auf den folgenden qPCR-Reaktionsparametern bewertet: Ct-Mittelwert, Ct-SD, CV, Schmelzkurvenanalyse (Daten nicht vorgestellt).