Purification of Amplifiable Nucleic Acid from Oil Palm and Rice Leaves and Seeds Using the Maxwell® 16 LEV Plant DNA Kit

Purification of Amplifiable Nucleic Acid from Oil Palm and Rice Leaves and Seeds Using the Maxwell® 16 LEV Plant DNA Kit

12 Jan 2016

This poster evaluates a novel cellulose-based paramagnetic particle technology to purify nucleic acid (NA) from Elaeis guineensis (oil palm) and Oryza sativa (rice) plant leaf and seed lysates in an automated format using the Maxwell® 16 System. Plant NA purification provides a variety of unique challenges due to the broad diversity of plants, their extracellular structures, and endogenous compounds (e.g. polysaccharides, phenolics) that can co-purify with NA and inhibit downstream enzymatic assays. Extracted NA were evaluated for yield and amplifiability in qPCR and RT-qPCR, and the data collected supports the broad utility of the novel cellulose-based technology and Maxwell® 16 System for automated plant NA purification.

GoTaq® 2-Step RT-qPCR System

Promega Corp.

GoTaq®2-Step RT-qPCR System is a reagent system for quantitative analysis of RNA using a two-step reverse transcription-quantitative PCR (RT-qPCR) method. The first step consists of cDNA synthesis using the GoScript™ Reverse Transcription System. Optimized reagents and ultra-active, GoScript™ Reverse Transcriptase result in high-efficiency, full-length cDNA synthesis from rare and abundant targets with up to 5 µg of RNA input. In the second step, cDNA is quantified using the GoTaq®qPCR Master Mix. Formulated with a hot-start DNA polymerase, the GoTaq® qPCR Master Mix is optimized for reproducibly specific, sensitive, highly efficient qPCR using the standard or fast mode of a real-time PCR instrument. The GoTaq® qPCR Master Mix contains BRYT™ GREEN dye which can be twice as bright as of SYBR® Green I. The superior performance of these two steps allows researchers to observe earlier quantification cycle values over broader linear ranges. The GoTaq® 2-Step RT-qPCR System provides a ready-to-use kit for analyzing a wide range of RNA targets by combining the high-activity of GoScript™ Reverse Transcription System with the ultra-bright fluorescence of GoTaq® qPCR Master Mix. The system is robust, requires little optimization, is flexible for a variety of applications, and carries the PCR performance guarantee.

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Maxwell® 16 LEV RNA FFPE Purification Kit

Promega Corp.

  The Maxwell® 16 LEV RNA FFPE Purification Kit is designed to extract RNA rapidly from FFPE samples using the Maxwell® 16 Instrument (Cat.# AS2000 and AS3000).   The FFPE kit is specifically designed for optimal purification of RNA from 5μm thin sections of formalin-fixed, paraffin-embedded (FFPE) tissue samples. The Maxwell® 16 LEV RNA FFPE Purification Kit is optimized to yield a final RNA concentration that maximizes RNA yield, eliminating the need to concentrate the extract prior to amplification. Lysates are placed directly into the cartridges, and RNA ready for amplification is obtained in approximately 60 minutes (after proteinase K digest). This kit does not use hazardous xylene, thereby providing a much safer method than many competitive kits. Additionally, quality testing demonstrates virtually no PCR inhibitors in purified samples. Features & Benefits: Automated Platform for Extracting RNA from FFPE Mammalian Tissues: Provides an alternative to manual competitive offerings. Walkaway Automated Extraction from up to 16 Samples in a Single Run: Provides superior ease of use over competition. Provides High Yield, Pure and Amplifiable RNA from FFPE Mammalian Tissue Samples: Minimizes sample waste and reruns. Simple Method with No Organic Solvents for Deparaffinization: No exposure to toxic organic reagents to deparaffinize the FFPE tissue sample prior to RNA purification. Decreased Processing Time: Protocol can be completed within a single 8-hour shift. Applications: Automated RNA purification from FFPE samples.

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QuantiFluor® ONE dsDNA Dye

Promega Corp.

  The QuantiFluor® ONE dsDNA Dye is a fluorescent double-stranded DNA-binding dye (504nmEx/531nmEm) prepared for an “add-and-read” format making sample quantitation easy. This dye enables sensitive quantitation of small amounts of double-stranded DNA (dsDNA).   The QuantiFluor® ONE dsDNA Dye is intended for use as a general purpose reagent in the US and is suitable for use by clinical laboratories as a component of laboratory developed tests. Outside of the US, this product is intended for research use only. The QuantiFluor® ONE dsDNA Dye can be used with any fluorometer that is capable of measuring fluorescence at the appropriate excitation and emission wavelengths. QuantiFluor® ONE dsDNA Dye Features & Benefits: Easy to Use - Add-and-read format makes this dye simple to use—no dilutions, no extra tubes. Sensitive - Significantly better sensitivity compared to absorbance at 260nm (NanoDrop®), allowing users to quantitate their low-concentration samples with confidence. Highly Specific to dsDNA - Minimal binding to ssDNA, RNA, protein and interfering compounds. Flexible with Instrumentation - Compatible with any fluorometer capable of measuring the appropriate fluorescence excitation and emission spectra. Applications: dsDNA quantitation.

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