Overview

• Purify sequencing extension products from dye terminators, primers and other contaminants
• Also purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations.
• High recovery
• Fast and efficient spin column format
• Also available in 96 well format for high throughput

This kit provides a rapid spin column procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on spin-column chromatography without the use of phenol, chloroform or alcohol precipitation. The kit provides a high quality product with up to 90% recovery.

The kit is also available in a 96-well format for high-throughput sequencing reaction clean-up.  Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.

Details

Supporting Data

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Kit Specifications – 96-well
Binding Capacity Per Well	15 μg
Maximum Loading Volume Per Well	500 μL
Average DNA Recovery	Up to 90%
Time to Complete 96 Purifications	20 minutes
Minimum Elution Volume	50 μL

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 34500 (50 preps)	Cat. 34400 (2 x 96-well plate)
Binding Buffer C	30 mL	3 x 30 mL
Wash Solution A	12 mL	2 x 20 mL
Elution Buffer B	8 mL	2 x 15 mL
Spin Columns	50	–
Collection Tubes	50	–
Elution Tubes (1.7 mL)	50	–
96-Well Plate	–	2
Adhesive Tape	–	4
96-Well Collection Plate	–	2
96-Well Elution Plate	–	2
Product Insert	1	1

Propargyl-PEG9-bromide consists of a propargyl group and a bromide group. The propargyl group reacts with azide compounds in copper catalyzed Click Chemistry reactions. The bromide (Br)can be used as a leaving group for nucleophilic substitution reactions. The 9-unit PEG spacer improves the hydrophilicity of the molecule. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG9-bromide consists of a propargyl group and a bromide group. The propargyl group reacts with azide compounds in copper catalyzed Click Chemistry reactions. The bromide (Br)can be used as a leaving group for nucleophilic substitution reactions. The 9-unit PEG spacer improves the hydrophilicity of the molecule. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Introduction

This kit is used for extracting total pathongen nucleic acid from low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate, culture, etc. The purified DNA/RNA is ready for downstream clinical in vitro detection such as Real Time PCR, biochip analysis, NGS and other related detection.

Details

Specifications

Features	Specifications
Main Functions	Extract pathogen DNA/RNA from whole blood, body fluid, serum, plasma, sputum, immersion solution, tissue homogenate supernatant, etc.
Applications	RT-PCR，PCR，NGS
Products	Pathogen DNA and RNA
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Whole blood, plasma, serum, immersion solution, tissue homogenate supernatant
Sample amount	200μl
Elution volume	≥30μl
Time per run	30 mins
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA/RNA is released into the lysate. Transfer to an adsorption plate and filter column. DNA/RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA / RNA is finally eluted with low-salt buffer (10 MmTris, pH 8.0).

Advantages

• Fast – column purification, several samples can be finished in 30 mins
• 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	IVD4179
Purification Times	50 Preps
HiPure Viral Mini Column	50
2ml Collection Tubes	100
2ml Bead Tubes	50
Protease K	50 mg
Protease Dissolve Buffer	5 ml
DNase I (Powder)	10 mg
DNase Buffer	6 ml
Buffer CLB	100 ml
Buffer SDS	5 ml
Reagent DX	1.5 ml
Buffer ACL	30 ml
Buffer VHB*	22 ml
Buffer RW2*	20 ml
Buffer AVE	15 ml

Storage and Stability

Proteinase K and DNase I (Powder) should be stored at 2–8°C upon arrival. However, short-term storage (DNase I up to 1 week, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affecttheir 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 used for extracting total pathongen nucleic acid from low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate, culture, etc. The purified DNA/RNA is ready for downstream clinical in vitro detection such as Real Time PCR, biochip analysis, NGS and other related detection.