The Multiplexing Index Primers contain primer mix for multiplexing library samples for Next Generation Sequencing (NGS) on the illumina platform. Multiplexing of NGS library samples will reduce sequencing costs by pooling multiple NGS libraries into a single flow cell lane.

With library multiplexing, unique index sequence is added to individual sample during NGS library preparation.  Therefore, each DNA molecule can be identified after pooling of multiple samples based on the index information they have.

Each of our index primers contains a unique index sequence with 6 bases that can be used to identify libraries. Library multiplexing up to 48 samples is possible.

Multiplexing Index Primers (illumina platform): Even distribution of 48 samples using index primers. 48 libraries were made using the BioDynami NGS DNA Library Prep Kit (Cat. # 30009) and the BioDynami Multiplexing Index Primers (Cat. # 30072). Libraries were pooled at equal concentration and sequenced on the illumina HiSeq 2500. The numbers of reads from 48 libraries were analyzed.

List of index sequence for the primers (each of the index primer mix contains universal primer and one of the index primers). Index number and the index sequence are listed.

List of indexes can be downloaded Here.

Sequence of the final library with index location:

The Multiplexing Index Primers contain primer mix for multiplexing library samples for Next Generation Sequencing (NGS) on the illumina platform. Multiplexing of NGS library samples will reduce sequencing costs by pooling multiple NGS libraries into a single flow cell lane.

Overview

• Detection kit for Toxoplasma gondii
• Available in TaqMan format for analysis

Toxoplasma gondii is a single-celled organism which causes Toxoplasmosis, the most common parasitic infection worldwide affecting animals as well as humans. Cats are the only animal in which sexual reproduction of the organism occurs, and for this reason cats are the only domestic animal which has the potential to shed T. gondii eggs. When disease does occur, it may develop following primary infection due to an inadequate immune response to stop the invasive tachyzoites or as a result of a reactivated infection due to a compromised immune system. The most common symptoms of the disease are anorexia, weight loss, lethargy, dyspnea, ocular symptoms and pyrexia. As toxoplasmosis can be transmitted to humans it represents a serious health risk for people living in close contact with infected animals. Infection is especially dangerous for people with suppressed immune systems and cancer patients as well as pregnant women.

Toxoplasma gondii TaqMan PCR Kit,
100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

Toxoplasma gondii TaqMan PCR Probe/Primer Set and Controls,
100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s PCR Master Mix (#28007) or customer supplied master mix

Details

Supporting Data

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Storage Conditions

All kit components can be stored for 2 years after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.

Component	TaqMan Probe Cat. TM44750 (100 preps)	TaqMan Probe Cat. TM44710 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 µL	–
Toxoplasma gondii Primer Probe Mix	280 µL	280 µL
Toxoplasma gondii Positive Control	150 µL	150 µL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Introduction

The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation miRNA from  0.3-0.5ml serum or plasma using magnetic particles
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	Serum, plasma, acellular samples
Sample amount	300μl

Principle

This product is based on the purification method of high binding magnetic particles. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After adding magnetic particles and binding solution, RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption.The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally RNA was eluted by Elution Buffer.

Advantages

• Safe – no phenol chloroform extraction and no use of Trizol reagent
• Fast – several samples can be extracted in 60 minutes by column method

Kit Contents

Contents	R662801	R662802	R662803
Purification Times	48 Preps	96 Preps	5 x 96 Preps
MagPure Particles N	1.7 ml	3.5 ml	17 ml
Buffer CFL	6 ml	12 ml	60 ml
Buffer CPL	1.8 ml	3.5 ml	20 ml
Buffer MGW1*	30 ml	60 ml	250 ml
Buffer MW2*	12 ml	25 ml	100 ml
RNase Free Water	10 ml	20 ml	60 ml

Storage and Stability

MagPure Particle N should be stored at 2–8°C upon arrival. However, short-term storage (up to 8 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 kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.