Introduction

This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.

Details

Specifications

Features	Specifications
Main Functions	Extract Pathogen RNA/DNA from 0.5ml plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution for mNGS application, remove host background nucleic acid.
Applications	RT-PCR，Real Time PCR, biochip analysis, NGS
Products	Pathogen DNA / RNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	low/cell-free samples such as plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution
Sample amount	0.5 ml
Adaptive instrument	Nucleic acid extractor, pipetting workstation

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. After adding magnetic particles and binding solution, DNA/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 DNA/RNA was eluted by Buffer NFW.

Kit Contents

Contents	R667200B	R6672-02B
Purification Times	24 Preps	96 Preps
2ml Bead Tubes	24	96
Proteinase K	12 mg	50 mg
Protease Dissolve Buffer	1.8 ml	3 ml
Buffer SDS (20%)	1.8 ml	8 ml
MagBind Particles	0.6 ml	2.5 ml
Buffer MLB	15 ml	60 ml
Buffer MW1*	13 ml	44 ml
Buffer MW2*	6 ml	50 ml
Buffer AVE	5 ml	30 ml

Storage and Stability

MagBind Particles and Proteinase K Solution 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 18 months under these conditions.

This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.

Overview

• Reliable and cost-effective
• Non-invasive, user-friendly sample collection and preservation in one convenient kit
• Preserved DNA is stable for 2 years at ambient temperature
• Compatible with any commercial DNA isolation methods including Norgen’s Saliva DNA Isolation Kits (Cat. RU45400)
• Samples are non-infectious and can be handled and shipped safely
• Shipping accessories can be purchased separately
• High quality DNA is suitable for sensitive downstream applications including PCR, qPCR, sequencing, SNP analysis, microarrays, RFLP and Southern Blot Analysis

Norgen’s Saliva DNA Collection and Preservation Devices are designed for 1) simple and non-invasive saliva collection and 2) preservation of DNA in saliva samples at ambient temperature. Each of the 50 Saliva DNA Collection and Preservation Devices consists of 3 components:

• Saliva Collection Funnel and Collection Tube
• Collection Tube Cap
• Norgen’s Saliva DNA Preservative contained within a sealed squeezable ampoule

Saliva samples are collected by spitting inside the Collection Funnel which has been assembled with the Collection Tube. After collecting the required volume of saliva the Collection Funnel is removed and the contents of the Preservative Ampoule are then added and mixed with the collected saliva. The Saliva Collection Tube is subsequently sent to the laboratory for DNA isolation and analysis. DNA can be isolated from the preserved saliva samples using Norgen’s Saliva DNA Isolation Kit (Cat. RU45400). Each of Norgen’s Collection Tubes is labeled with a unique serial number that can be used for secure and anonymous tracking of the sample. The saliva DNA in preserved samples is stable for more than 2 years at room temperature. This kit is ideal for collecting and preserving DNA samples for epidemiological and population studies.

Saliva DNA Preservative

Norgen’s Saliva DNA Preservative is an aqueous storage buffer designed for rapid cellular lysis and subsequent preservation of DNA from fresh specimens. The buffer prevents the growth of Gram-negative and Gram-positive bacteria and fungi, and also inactivates viruses allowing the resulting non-infectious samples to be handled and shipped safely. In addition, the buffer eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperature. The components of the buffer allow samples to be stored for more than 2 years without any detectable DNA degradation.

DNA Isolation from Preservative

Prior to saliva DNA isolation, vortex the Collection Tube containing preserved saliva for 10 seconds and incubate at 55°C for one hour. Saliva DNA can now be isolated from the preserved saliva samples using any commercially available method, including Norgen’s Saliva DNA Isolation Kit (Cat. RU45400), Norgen’s Saliva DNA Isolation 96-Well Kit (Cat. RU35200) and Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System) (Cat. RU55400, RU62900). Also, Norgen’s Saliva DNA Isolation Reagent Kit (up to 4 mL) (Cat. RU35720) allows for DNA isolation from the preserved saliva samples using an alcohol precipitation method. The purified DNA is of the highest quality and can be used in a number of downstream applications including PCR, NGS and microarray analysis.

Details

Supporting Data

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Kit Specifications
Volume of Saliva Collected	2 mL
Volume of Saliva-Preservative Mix	4 mL
Preservation Temperature	Room Temperature
Preservation time	Over 2 years at room temperature

Shelf Life and Handling

• When stored at room temperature, unused Norgen Saliva Preservative Tubes are stable through to the collect-before date without any reduction in performance. Please see the device insert or tube label for collect-before date.
• Once collected, saliva is stable for more than 2 years when kept tightly sealed and stored at room temperature.
• The Collection Tube, the Collection Funnel and the Device Container of each Individual Saliva DNA Collection and Preservation Device are recyclable.

Kit Components	Cat. RU49000 (50 Devices)
Individual Saliva DNA Collection and Preservation Devices	50
Donor Procedure Flowchart	1
Product Insert	1
Individual Saliva DNA Collection and Preservation Device Contents
Saliva Collection Funnel and Collection Tube	1
Collection Tube Cap	1
Preservative Ampoule	1
Donor Instructions	1

The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.

NGS Single Stranded DNA Library Prep Kit workflow

Three index types are available for the kit:

Non-index (Cat.# 30081): Libraries do not have index.

Index(Cat.# 30082): Each of the index primers has a unique 6-base index sequence that can be used to identify libraries. ssDNA library multiplexing is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30083): ssDNA sample multiplexing up to 96 libraries is possible with unique dual indexes. We have developed a Four-Base Difference Index System. This makes it possible to generate indexes with at least 4 bases different from each other in the 8 bases index length. The unique dual indexing primers remove NGS errors (example: de-multiplexing errors, read mis-assignment, index hopping etc). Index information can be downloaded here.

Kit advantages:

• • • Quick and simple Protocol
      • Quick: Total protocol time is only 1.5 hours
      • Simple: Hands-on time only 10 minutes
    • Easy procedure based on:
      • The ready-to-use master mix: Made reaction setup easy
      • Less reaction components: Simplify the reaction preparation
    • Less magnetic beads needed: Reduced more than 50% of the beads for cleanup steps
    • Directional library
    • Single stranded DNA as input: From 50 ng to 500 ng

NGS Single Stranded DNA Library Prep Kit has similar library conversion efficiency and yield as compared to a regular DNA library prep kit.

Alignment rate and duplication rate: comparison of single stranded DNA library kit versus double stranded DNA library prep kit.

The NGS Single Stranded DNA Library Prep Kit (illumina platform) was developed for construction of high quality libraries using sheared single stranded DNA (50 ng – 500 ng) as input. The kit is ideal for making NGS libraries with samples of denatured DNA, viral DNA, highly degraded ancient DNA and other single stranded DNA. The workflow of the ssDNA kit is simple: make the libraries in two steps followed by PCR and cleanup steps. The libraries will be ready in just 1.5 hours with a 10 minutes of hands-on time. The library pooling is possible dependent on the type of index. The final library is strand specific.