Overview

• Detection kits for the HBV
• Available in TaqMan format for analysis

The Hepatitis B virus (HBV) is mainly transmitted via blood or blood products. In addition, sexual, oral and perinatal infections are also possible. Early symptoms of the infection include appetite loss, vomiting and abdominal symptoms, with approximately 10-20% of those patients developing fever as well as rheumatoid joint and muscle pain. Jaundice, which may be accompanied by itching, will then develop within 2-14 days. Fulminant hepatitis then occurs in about 1% of all infected patients, which in severe cases may be fatal. Of those individuals infected by HBV, 5-10% will develop chronic liver inflammation which may progress to cirrhosis of the liver or, in the worst case, primary liver cell carcinoma. The HBV virus is a hepadnavirus that has a circular genome composed of partially double-stranded DNA.

HBV 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

HBV 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

For research use only and NOT intended for in vitro diagnostics.

Details

Supporting Data

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Storage Conditions and Product Stability
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	Cat. TM29250 (100 rxns)	Cat. TM29210 (100 rxns)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
HBV Primer & Probe Mix	280 μL	280 μL
HBV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• ​Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment, nor carrier RNA
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents, nor overnight incubation required
• Concentrate isolated exosomal RNA and are free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 50 µL to 1 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 1 mL to 4 mL.

Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 4 mL to 10 mL.

Details

Supporting Data

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Kit Specifications
Minimum Plasma Input	4 mL
Maximum Plasma Input	10 mL
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50 – 100 µL
Time to Complete 10 Purifications	40 – 45 minutes
Average Yields*	Variable depending on specimen

*Please check page 5 of the product insert for the average yields and the 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.

Important Note
This kit is suitable for the purification of exosomes 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.

Component	Cat. 59500 (50 preps)	Cat. 59600 (25 preps)	Cat. 59700 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 200μl whole blood
Applications	PCR, southern bolt and virus detection, etc
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount	200μl
Elution volume	≥50μl
Time per run	≤60 minutes

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. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA 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 was eluted by Elution Buffer.

Advantages

• High binding force – suitable for handling DNA rich samples, such as whole blood, buffy coat, concentrated blood, etc.
• Fast – polydisperse magnetic beads, fast magnetic response and short extraction time
• High purity – the obtained DNA can be directly used for second-generation sequencing, PCR based detection, gene bank, etc.
• Automatic – saving time andlabor and safer

Kit Contents

Contents	D631101	D631102	D631103
Purification Times	48	96	480
MagPure Particles	1.2 ml	2.5 ml	11 ml
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	1.8 ml	5 ml	15 ml
Buffer AL	15 ml	30 ml	120 ml
Buffer BD	5 ml	10 ml	50 ml
Buffer GW1	22 ml	53 ml	220 ml
Elution Buffer	15 ml	30 ml	100 ml

Storage and Stability

Proteinase K, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage(up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry 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 this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.