PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(endo-BCN-PEG2-ethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The DBCO groups are commonly used for copper-free Click Chemistry reactions due to their strain promoted high energy. The hydrophilic PEG chain allows for increased water solubility.

PEG3-(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)-(Amino-Tri-(endo-BCN-PEG2-ethoxymethyl)-methane) is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The DBCO groups are commonly used for copper-free Click Chemistry reactions due to their strain promoted high energy. The hydrophilic PEG chain allows for increased water solubility.

Introduction

This product is suitable for extracting total viral nucleic acid from cell-free/low-content cell biological samples such as body fluids, serums, plasma, soaking solutions, tissue homogenate supernatant, and culture supernatant. The Purified DNA/RNA is used for RT-PCR and PCR detection.

Details

Specifications

Features	Specifications
Main Functions	Extract viral DNA/RNA from200μl samples by magnetic beads
Applications	RT-PCR，PCR，NGS
Products	Viral DNA / RNA, body cell DNA / RNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technolog
Process method	Manual or automatic
Sample type
Sample amount	200μl
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Yield
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

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/RNA is released into the lysate. 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 Nuclease Free Water.

Advantages

• Fast – several samples can be extracted in 40 minutes by column method
• High quality – high purity total RNA / DNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – DNA/RNA can be recovered at the level of PG

Kit Contents

Contents	IVD5412
Purification Times	200 Preps
MagPure Particles N	5 ml
PK/Carrier RNA	50 mg
Protease Dissolve Buffer Blue	5 ml
Buffer MLB	120 ml
Buffer MW1	53 ml
RNase Free Water	30 ml

Storage and Stability

This kit is shipped and stored at room temperature and is valid for 12 months.

Experiment Data

This product is suitable for extracting total viral nucleic acid from cell-free/low-content cell biological samples such as body fluids, serums, plasma, soaking solutions, tissue homogenate supernatant, and culture supernatant. The Purified DNA/RNA is used for RT-PCR and PCR detection.

Overview

• Optimized for low input RNA, especially from bodily fluids such as plasma or serum at 1ng of RNA
• Simple and quick workflow: library could be prepared in less than 5 hours
• No gel purification for selected types of samples
• Protocol optimized for RNA isolated from different types of input, including liquid biopsies (blood, plasma, serum and urine)
• Complements Norgen’s Best-in-Class Total RNA (including microRNA) Purification Technology

The Small RNA Library Prep Kit for Illumina consists of all the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform. All molecular reagents including adaptors, primers, enzyme mixes and buffers are provided. A purification module is also provided for rapid purification of nucleic acid products generated at various steps of the workflow. The purification module utilizes Norgen’s patent resin technology which enhances recovery of desired library intermediates or final products. The library prep workflow could be used for different forms of input including purified total RNA or enriched small RNA, as well as RNA from low content inputs such as plasma, serum and urine.

Workflow

Details

Supporting Data

Figure 1 / 4

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Kit Specifications
Number of preps	24
Number of indices provided	12 (for 24 preps, indexes 1-24 or 25-48)
Time required to complete 6 libraries	As little as 5 hours
Input RNA required	As little as 50 pg

Storage Conditions and Product Stability
Some components require storage at -20°C, 4°C or room temperature. See individual components and box labels for storage conditions.

Step	Component	Cat. 64600 (24 preps)
3′ AdaptorLigation to Template RNA	3′ Adaptor	30 µL
	3′ Adaptor Ligation Master Mix	320 µL
	T4 RNA Ligase 2 (Truncated)	35 µL
5′ Adaptor Ligation	5′ Adaptor	30 µL
	5′ Adaptor Ligation Master Mix	320 µL
	T4 RNA Ligase 1	35 µL
cDNA Synthesis from Ligated RNA Product	Reverse Primer	30 µL
	cDNA Synthesis Master Mix	220 µL
	TruScript ReverseTranscriptase	35 µL
PCR Amplification	2x NGS PCR Master Mix	1.32 µL
	PCR Reverse Primer	81 µL
	Forward Index Primer	Included in Small RNA Library Prep Forward Index Primers (# 64640 or # 64610)
Size Selection	NGS MW Ladder	50 µL
	NGS Control Ladder	50 µL
	Loading Dye	300 µL
	Nuclease-Free Water	1.25 µL

Component	Cat. 63500 (75 preps)
Buffer RL	40 mL
Wash Solution A	38 mL (Reconstituted to 128 mL)
Elution Solution A	6 mL
Columns	75
Gel Filtration Columns	24
Collection Tubes	75
Elution Tubes	75
Product Insert	1