Everything you need to run a trial PACE 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.

About

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
• PCR-grade water

qPCR machine or Thermocycler + Fluorescent plate reader

PCR plate or equivalent and appropriate optically clear seal

PCR-grade water

• Format: 96-well microtiter plate (12 test strips of 8 wells)
• Standards: 0 | 0.03 | 0.10 | 0.2 | 2 ppb
• Incubation Time: 45 Minutes
• Compatible for use with US EPA Method 546

Description

The Cylindrospermopsin plate kit is a competitive enzyme-labeled immunoassay. The Cylindrospermopsin sample extract and calibrators are pipetted into the test wells followed by the Cylindrospermopsin antibody into the test wells to initiate the reaction. During the 30 minutes incubation period, Cylin-drospermopsin from the sample and Cylindrospermopsin antigen compete for binding to the Cylindrosper-mopsin antibody. The Cylindrospermopsin antibody is captured on the walls of the test well. Following this 30-minute incubation, the contents of the wells are removed and the wells are washed to remove any unbound Cylindrospermopsin and free Cylindrospermopsin antibody. After wash, 1X HRP-conjugated Antibody#2 is added for 30 minutes incubation. The wells are washed afterwards, and a clear substrate is then added to the wells and any bound enzyme conjugate causes the conversion to a blue color. Following a 15-minute incubation, the reaction is stopped and the amount of color in each well is read. The color of the unknown samples is compared to the color of the calibrators and the Cylindrospermopsin concentration of the samples is derived.

Format:

• • 96-well microtiter plate (12 test strips of 8 wells)
• • Standards: 0 | 0.03 | 0.10 | 0.2| 2 ppb
• • Incubation Time: 45 Minutes
• Compatible for use with US EPA Method 546

EPA 10-Day drinking water Health Advisories for Cylindrospermopsin: 

Do not Drink – 0.7 μg/L for bottle fed infants and preschool children, pregnant and nursing woman, elderly immunocompromised and liver conditions.

Do not Drink – 3.0 μg/L for school age children to adults.

Do Not Use – 20 μg/L

EPA Draft Human Health Recreational Ambient Water Quality Criteria to protect human health: 8 μg/L.

Format: 96-well microtiter plate (12 test strips of 8 wells)
Standards: 0 | 0.03 | 0.10 | 0.2 | 2 ppb
Incubation Time: 45 Minutes
Compatible for use with US EPA Method 546

Introduction

HiPure Insect DNA Kits provides a simple and rapid solution for total DNA extraction of insect tissue samples. This kit is based on silica gel column purification technology without toxic phenol chloroform extraction and time-consuming alcohol precipitation. The whole extraction process only takes 30 minutes. HiPure Insect DNA Kit can process tissue samples less than 10mg at a time. Hipure Insect DNA 96 kit can process 96 insect tissue samples at a high throughput. The obtained DNA can be directly used in PCR, Southern blot, viral DNA detection and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from <10 mg insect tissue
Applications	PCR, southern bolt and virus detection, etc
Purification method	Midi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Insect tissue samples
Sample amount	<10 mg
Elution volume	≥15μl
Time per run	≤30 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principles

This product is based on silica column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High purity DNA – can be used in sensitive downstream applications such as multiplex and quantitative pcr
• Fast – several samples can be extracted in less than 30 minutes
• Good repeatability – suitable for extracting high-yield DNA from insect tissue samples
• Safe – without phenol chloroform extraction and alcohol precipitation

Kit Contents

Contents	D312902	D312903
Purification Times	50	250
HiPure DNA Mini Columns I	50	250
2ml Collection Tubes	50	250
Buffer ITL	30 ml	120 ml
Buffer IL*	30 ml	120 ml
Buffer GW1*	22 ml	110 ml
Buffer GW2*	20 ml	2 x 50 ml
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	15 ml
Buffer AE	15 ml	60 ml

Storage and Stability

Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in thiscase buffers should be redissolved before use. Make sure that all buffers areat room temperature when used.

HiPure Insect DNA Kits provides a simple and rapid solution for total DNA extraction of insect tissue samples. This kit is based on silica gel column purification technology without toxic phenol chloroform extraction and time-consuming alcohol precipitation. The whole extraction process only takes 30 minutes. HiPure Insect DNA Kit can process tissue samples less than 10mg at a time. Hipure Insect DNA 96 kit can process 96 insect tissue samples at a high throughput. The obtained DNA can be directly used in PCR, Southern blot, viral DNA detection and other experiments.