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.

OverView

T4 DNA Ligase is an ATP and Mg2+ dependent dsDNA ligase which catalyses the formation of a phosphodiester bond between 3’-hydroxyl and 5’-phosphate termini in duplex DNA, duplex RNA and some DNA/RNA hybrids. T4 DNA Ligase is active on both blunt-end and cohesive-end substrates. It is also completely inactivated by incubating at 70°C for 10 minutes.

This is a high-quality (commercial grade) version of the T4 DNA Ligase. T4 DNA Ligase is recombinantly produced in E. coli. ArcticZymes’ T4 DNA Ligase is extensively tested for contaminating DNase and RNase activities as well as residual host-cell gDNA.

Key Features

• ATP and Mg2+ dependent dsDNA ligase
• Easily heat-inactivated at 70°C for 10 minutes
• Extensively tested for contaminating DNase and RNase activities as well
  as residual host-cell gDNA

Applications

• Ligation of dsDNA
• NGS library prep
• Molecular cloning

Figures

T4 DNA Ligase is an ATP and Mg2+ dependent dsDNA ligase which catalyses the formation of a phosphodiester bond between 3’-hydroxyl and 5’-phosphate termini in duplex DNA, duplex RNA and some DNA/RNA hybrids. T4 DNA Ligase is active on both blunt-end and cohesive-end substrates. It is also completely inactivated by incubating at 70°C for 10 minutes.

Introduction

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA and miRNA from cell and tissue without MagZol reagent
Applications	RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Cell, Animal tissue, Plant tissue
Sample amount	Cells: ≤ 5 x 10^6, Animal tissue:<10mg
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Biological samples are first lysed and homogenized in a highly denaturing guanidine isothiocyanate-containing buffer, which immediately inactivates DNases and RNases to ensure isolation of intact DNA and RNA. The lysate is then passed through a Mini spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. Flow-through from the column is digested by Proteinase K in the presence of ethanol. This optimized digestion, together with the subsequent addition of further ethanol, allows appropriate binding of total RNA, including miRNA, to the column. Contaminants are efficiently washed away and high-quality RNA is eluted.

Advantages

• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 40 minutes
• High applicability – samples including animals, plants, bacteria, cells, etc.
• High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
• Safe – no phenol chloroform extraction and no use of Trizol reagent

Kit Contents

Contents	R431102	R431103
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
Proteinase K	48 mg	240 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer RLC	40 ml	200 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	60 ml

Storage and Stability

Proteinase K 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 their 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.

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.