Introduction

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from tissue / blood / body fluid / swab /dry blood spots
Applications	PCR, qPCR, southern bolt and virus detection, etc
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Tissue, cell, blood, saliva, swab, blood spot,semen and other clinical samples
Sample amount	Solid tissue: 1-10mg; Anticoagulant blood: 200μl
Elution volume	≥20μl
Time per run	30 – 60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
• Fast – without separation of leukocytes, organic extraction or ethanol precipitation
• Simple – all nucleic acids can be obtained by direct digestion
• Wide applicability- handle a variety of liquid samples

Kit Contents

Contents	D301802	D301803
Purification Times	50	250
HiPure DNA Mini Columns I	50	250
2ml Collection Tubes	100	500
Buffer ATL	30 ml	150 ml
Buffer AL	30 ml	150 ml
Buffer GW1	22 ml	88 ml
Buffer GW2	12 ml	50 ml
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	15 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 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.

Experiment Data

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Overview

• Rapid isolation of both small and large species of DNA from urine
• Convenient spin column format
• Effective removal of PCR inhibitors
• Purified DNA is highly suited to sensitive downstream applications
• Allows for the purification of viral DNA from urine

Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.

This kit is fully compatible with Norgen’s Urine Collection and Preservation Tubes.

Urine DNA Isolation Kit

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.

Urine DNA Isolation Kit (Slurry Format)

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.

Urine DNA Isolation Maxi Kit (Slurry Format)

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.

Background

DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.

Details

Supporting Data

Figure 1 / 8

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Kit Specifications
Volume of Urine Processed	1.75 mL
Average Yield*	Up to 50 ng
Size of DNA Purified	Large (> 1 kb) and small (150-250 bp)
Time to Complete 10 Purifications	30 minutes hands-on time (plus a 1 hour incubation)

* Yield will vary depending on the type of sample processed


Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.

Component	Cat. 18100 (50 preps)	Cat. 48800 (50 preps)	Cat. 50100 (50 preps)
Binding Solution K	15 mL	–	–
Slurry B1	–	18 mL	18 mL
Proteinase K in Storage Buffer	2 mL	–	–
Pronase K in Storage Buffer	2 mL	–	–
Soluton WN	9 mL	–	–
Binding Buffer A	–	–	50 mL
Lysis Buffer A	–	30 mL	30 mL
Wash Solution A	–	38 mL	38 mL
Wash Solution B	30 mL	–	–
Wash Solution D	9 mL	–	–
Binding Solution K	15 mL	–	–
Elution Buffer B	15 mL	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Filter Spin Columns	–	50	50
Collection Tubes	50	50	50
Elution Tubes (1.7 mL)	100	100	100
Product Insert	1	1	1