Biotin-PEG3-alkyne

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

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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

Introduction

Magnetic bead nucleic acid purification technology uses nano or micron superparamagnetic material as the matrix, generally black ferric oxide or yellowish brown ferricoxide as the magnetic material. The surface of bead is coated with appropriate functional groups, which can adsorb nucleic acid. Magnetic beads commonly used for nucleic acids, containing carboxyl groups, hydroxyl groups, or silicon groups. Silicon-based magnetic beads are the most common, and its principle of adsorbing nucleic acid is consistent with the classical glass milk purification technology or glass fiber filter membrane purification method. Magpure particle is a kind of polydisperse fast speed silica magnetic beads. The core is ferricoxide, accounting for 50%, and the surface coating is silica, accounting for 50%. The product can be used for plasmid extraction, gel DNA recovery, product purification, genomic DNA and RNA extraction, and viral nucleic acid extraction.

Details

Specifications

Features	Specifications
Concentration	10 mg/ml
Appearance	Suspension of yellowish brown particles
Surface functional group	Carboxyl, COOH
Dispersibility	Monodisperse, spherical
Particle size	0.8-1 μm
Preservation conditions	Room temperature, valid for up to 2 years.It is recommended to store in 2-8°C to prevent microbial growth.
Magnetic response speed	120 seconds
Settling velocity	>2 hours
High salt mediated binding	No adsorption
Alcohol mediated binding	1M NaClO4/ethanol(50%), DNA/RNA recovery up to 90%
PEG8000 mediated binding	The recovery of DNA/RNA was up to 90%
DNase/RNase	Not detected
DNA residue	Not detected
Recommended application	Plasmid extraction, gel DNA recovery, genomic DNA extraction and RNA extraction.

Principle

Highsalt mediated binding: in the solution containing 2-4M guanidine isothiocyanate, Magpure particles can selectively recover DNA molecules, and impurities such as protein polysaccharides are not adsorbed.

Alcohol mediated binding: in the solution containing guanidine salt and alcohol (>25%), Magpure particles can selectively recover DNA/RNA molecules, and proteins and other impurities are not adsorbed.

After biological samples are treated with digestive solution or lysis Buffer, DNA/RNA is released from cells, organelles and protein complexes (ribosomes and nucleosomes) into reagents. After Magpure particles and binding solution are added, DNA/RNA is adsorbed to the surface of Magpure particles to form DNA/RNA bead complex. Under the action of the magnetic field, the magnetic beads are separated and collected, and the impurities such as protein are removed with the waste liquid. After two or three steps of further cleaning, the DNA/RNA magnetic bead complex is resuspended in sterilized water or TE buffer, and the DNA/RNA falls off from the surface of the magnetic beads, so as to achieve the purpose of purification.

Ordering information

CAT.No.	Product Name	Package
C14130	MagBind Particles	10 ml
C14131	MagBind Particles	100 ml

Purchase Guide

Features	MagPure Particles	MagPure Particles N	MagPure Particles G	MagPure Particles F	MagBind Particles
Cat.No.	C1410	C1411	C1412	C1414	C1413
Concentration	100mg/ml	70mg/ml	40mg/ml	50mg/ml	10mg/ml
Form	Amorphous and Porous	Amorphous and Porous	Porous	Amorphous	Nonporous
Surface function	Si-OH, Silica Beads	Si-OH, Silica Beads	Si-OH, Silica Beads	Si-OH, Silica Beads	COOH, Carboxyl Beads
Dispersion	Polydisperse	Polydisperse	Monodisperse	Monodisperse	Monodisperse
Particle Size	1.5-5μm	0.2-2μm	1-1.5μm	0.2-1.5μm	0.8-1μm
Color	Black	Yellowish Brown	Dark Brown	Dark Brown	Yellowish Brown
Magnetic response	15-30s	~60s	~30s	20s	120s
Settling Time (1ml)	>5min	>10min	>3min	>3min	>2h
Usage (0.2ml Sample)	20μl	20μl	20-30μl	20-30μl	20-30μl
DNA Recover Rate (only 4M GITC)	>80%	>80%	>80%	>80%	0
DNA Recover Rate (10% PEG8000/NaCl)	>85%	>85%	>85%	>85%	>90%
Recommended Use	gDNA/RNA Isolation from Blood, Tissue, Plant, Swab, Spots, Stool, Soil and etc.Viral DNA/RNA IsolationAgarose Gel DNA Purification	DNA/RNA Isolation from low nucleic acid content samplesPlasmid IsolationDNA/RNA Clean Up	Circulating DNA IsolationViral Nucleic acid IsolationgDNA Isolation FFPE DNA/RNA Isolation	Plasmid extractiongel DNA recoverygenomicDNA/RNA extraction viral nucleic acid extractionCirculating DNA extraction	DNA/RNA Clean Up and concentrationDNA/RNA Isolation from low nucleic acid content samplesResearch immuno assays

• The MagPure magnetic-particle technology combines the speed and efficiency of silica-based DNA purification with the convenient handling of magnetic particles. DNA binds to the silica surface of the magnetic particles in the presence of a chaotropic salt. DNA bound to the particles is then efficiently washed, considerably improving the purity of DNA. High-quality DNA is eluted. The automated purification procedure completely removes enzymes, nucleotides, and other contaminants and inhibitors. Purified DNA is suitable for direct use in downstream applications, such as sequencing and microarray analysis.

Magnetic bead nucleic acid purification technology uses nano or micron superparamagnetic material as the matrix, generally black ferric oxide or yellowish brown ferricoxide as the magnetic material. The surface of bead is coated with appropriate functional groups, which can adsorb nucleic acid. Magnetic beads commonly used for nucleic acids, containing carboxyl groups, hydroxyl groups, or silicon groups. Silicon-based magnetic beads are the most common, and its principle of adsorbing nucleic acid is consistent with the classical glass milk purification technology or glass fiber filter membrane purification method. Magpure particle is a kind of polydisperse fast speed silica magnetic beads. The core is ferricoxide, accounting for 50%, and the surface coating is silica, accounting for 50%. The product can be used for plasmid extraction, gel DNA recovery, product purification, genomic DNA and RNA extraction, and viral nucleic acid extraction.

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request