PC-Biotin-PEG4-PEG4-alkyne is useful for introducing a biotin moiety to azide-containing biomolecules using Cu(I)-catalyzed Click Chemistry. The extended hydrophilic spacer arm provides better solubility to the labeled molecules in aqueous media. Captured biomolecules can be efficiently photoreleased using near-UV, low intensity lamp (e.g. 365 nm lamp at 1-5 mW/cm2). Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

PC-Biotin-PEG4-PEG4-alkyne is useful for introducing a biotin moiety to azide-containing biomolecules using Cu(I)-catalyzed Click Chemistry. The extended hydrophilic spacer arm provides better solubility to the labeled molecules in aqueous media. Captured biomolecules can be efficiently photoreleased using near-UV, low intensity lamp (e.g. 365 nm lamp at 1-5 mW/cm2). Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Overview

• Concentrate circulating DNA into a flexible elution volume ranging from (25 µL – 50 µL)
• Isolate inhibitor-free cell-free circulating DNA
• Compatible with Streck Cell-Free DNA BCT® Tubes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
• Research use format is also available

These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.

Background

Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.

Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx

For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes

Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx

For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.

Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx

The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.

Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx

The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.

Details

Supporting Data

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Kit Specifications
Minimum Plasma/Serum Input	10 μL
Maximum Plasma/Serum Input	200 μL
Size of DNA Purified	≥ 50 bp
Elution Volume	25-50 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yields*	Variable depending on specimen

*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.

Component	Cat. Dx55100 (50 preps)	Cat. Dx55500 (50 preps)	Cat. Dx55600 (20 preps)	Cat. Dx55800 (10 preps)
Binding Buffer B	2 x 40 mL	40 mL	2 x 85 mL 1 x 12 mL	2 x 85 mL 1 x 40 mL
Proteinase K	2 x 1 mL	0.6 mL	6.5 mL	8 mL
Solution WN	18 mL	18 mL	18 mL	18 mL
Wash Solution A	38 mL	18 mL	18 mL	18 mL
Elution Buffer B	8 mL	8 mL	30 mL	30 mL
Micro Spin Columns	50	50	20	10
Mini Spin Columns	50	–	–	–
Midi Spin Columns	–	–	20	–
Maxi Spin Columns	–	–	–	10
Collection Tubes	100	50	20	10
Elution Tubes (1.7 mL)	100	50	20	10
Product Insert	1	1	1	1

Introduction

The Kit integrates phenol/guanidine-based lysis and silica membrane purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of samples and inhibit RNases. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable extracting up to 200μg total purified RNA from less than 100mg animal, plant, fungal samples,bacteria, cells and other samples.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA (include miRNA) from 100mg lipid tissue, tissue, cell, plant, body fluids using columns and MagZol reagent
Applications	RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Purification method	Mini spin column
Purification technology	Silica technology, acid phenol / guanidine extraction technology (MagZol pretreatment technology)
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissue, muscle fiber containing tissue,adipose tissue, cultured cells, lymphocytes, simple plants and other biological samples
Sample amount	Animal tissue sample: 1-60mg (spleen≤ 10mg)Cultured cells: 5 x 106 Plant leaves: 50-100mg
Yield	2-200μg
Elution volume	≥50μl
Time per run	≤30 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Tissue samples (10-100mg) are homogenized in MagZol Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30-100µl of RNase-free water.

Advantages

• Efficient DNA removal – one step RNA extraction can effectively remove genomic DNA
• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 30 minutes
• High applicability – samples including animals, plants, bacteria, cells, etc.
• High output – enable to process large samples and the yield can be up to 200μg

Kit Contents

Contents	R413002	D413003
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
MagZol Reagent	60 ml	270 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its 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.

Experiment Data

The Kit integrates phenol/guanidine-based lysis and silica membrane purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of samples and inhibit RNases. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable extracting up to 200μg total purified RNA from less than 100mg animal, plant, fungal samples,bacteria, cells and other samples.