Everything you need to run a trial PACE® allele-specific PCR Genotyping Reaction on your existing lab equipment. Each PACE Trial Kit includes Test DNA samples, PACE Genotyping Assays, PACE Master Mix and a comprehensive PACE Genotyping Trial Kit Manual.
WHO IS THIS TRIAL KIT FOR?
Anyone who wants to try PACE genotyping reagents in their lab for the first time with a set of validated DNA samples, SNP assays and PACE Master Mix.
TRIAL KIT OVERVIEW
Step 1. Dispense each of the three trial DNA samples (DNA 1, 2 and 3) plus water (No Template Control) in triplicate onto a PCR plate using the suggested volumes.
Step 2. Combine appropriate volumes of PACE Genotyping Master Mix with PACE Genotyping Assay in a tube, as directed, then mix.
Step 3. Dispense the combined mixtures into each of the wells containing DNA using volumes indicated. Each test now contains a complete PACE Genotyping Reaction.
Step 4. Seal your PCR plate with an optically clear seal and centrifuge to ensure all components are at the bottom of the wells.
Step 5.Thermally cycle the reaction plate using the thermal cycling conditions provided.
Step 6. Read the plate and compare data produced with the expected results provided in the manual. Simple!
PACE MECHANISM
More information on the PACE genotyping chemistry and how it works can be found here: www.3crbio.com/#pace. PACE allele-specific PCR is used for the detection of SNPs, Indels and other sequence variants.
REQUIRED COMPONENTS
qPCR machine or Thermocycler + Fluorescent plate reader
PCR plate or equivalent and appropriate optically clear seal
What is the genesig q16? The genesig q16 is a revolutionary instrument launched by Primerdesign Ltd. The instrument is designed to accompany the genesig Easy product range which includes kits for more than 550 different DNA testing applications. The genesig q16 is designed to make DNA testing affordable and easy for anyone in any business.
What is DNA testing? DNA testing is the most sensitive and precise way to detect and quantify the presence of a DNA target. The underlying technology within the genesig q16 is real-time quantitative PCR. The technology has been around for 20 years, but to date has been complex and expensive to perform. The genesig q16 changes all that.
What can I use it for? The genesig Easy product range includes tests for a massive range of applications:
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Incredibly easy to use Automated data analysis Robust, beautiful design Small and portable: 12cm footprint Over 550 kits available from a vast range of human, food and veterinary detection profiles Highly portable
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Background
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
EXTRAClean Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
EXTRAClean Plasma/Serum RNA Purification Midi Kit
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
EXTRAClean Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
All sizes, including miRNA and small RNA (<200 nt)
Average Yields¥
Variable depending on specimen
† This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR
¥ Please check page 6 for Average Plasma/Serum Yields and Common RNA Quantification Methods
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
