Introduction

HiPure RNA Clean Up Kit provides a rapid and easy method for the purification and concentrate RNA from enzymatic reactions or for desalting the RNA samples. RNA purified using HiPure RNA Clean Up Kit is ready for all downstream applications such as RT-PCR, Northern blotting, mRNA purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Purification and concentration of RNA（mRNA>200nt or miRNA）from transcription products, DNase cleavage products, labeled products, and crude RNA products
Applications	Sequencing, ligation, enzyme digestion, RT-PCR, labeling, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Crude RNA, enzymatic reaction solution
Sample amount	≤100μl
Recovery	90%
Elution volume	≥15μl
Time per run	≤15 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Binding buffer is added directly to the sample or other enzymatic reaction, and the mixture is applied to the column. Nucleic acids adsorb to the silica membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure RNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.

Advantages

• High recovery – recovery of RNA up to 90%
• Low elution volume – the elution volume can be as low as 15µl
• Fast – only 10 minutes for recovery by using column method
• General – suitable for various crude RNA products
• Wide range of fragment recovery:(>25nt RNA)

Kit Contents

Contents	R214402	R214403
Purification Times	50 Preps	250 Preps
Buffer GXP	30 ml	120 ml
Buffer RW2	20 ml	2 x 50 ml
RNase-Free Water	20 ml	60 ml
HiPure RNA Mini Columns I	50	250
2 ml Collection Tubes	50	250

Storage and Stability

The Kit 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.

HiPure RNA Clean Up Kit provides a rapid and easy method for the purification and concentrate RNA from enzymatic reactions or for desalting the RNA samples. RNA purified using HiPure RNA Clean Up Kit is ready for all downstream applications such as RT-PCR, Northern blotting, mRNA purification, nuclease protection, and in vitro translation.

Overview

• Ready-to-use
• Quantitative
• Highly Stable
• Precise
• Ten discrete fragments from 100 bp to 1000 bp
• Higher intensity reference band at 500 bp

The Norgen PCRSizer 100 bp DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our PCRSizer contains ten discrete fragments ranging from 100 bp to 1000 bp in 100 bp increments with a higher intensity reference band at 500 bp. This Ladder is ideal for PCR product size confirmation.

Contents:
1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).

Details

Supporting Data

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PCR Sizer 100 bp DNA Ladder (Cat# 11400) – 100 loads

Ladder Properties:
• Ten discrete bands, ranging from 100 bp to 1,000 bp
• Higher intensity band at 500 bp for easy reference

Fragment	Size (bp)	Mass (ng)
1	1000	78
2	900	70
3	800	62
4	700	55
5	600	47
6	500	78
7	400	31
8	300	24
9	200	31
10	100	24

Recommended Use:

Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.

Storage: 

Stable at room temperature. For longer term storage, -20°C is recommended.

This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.

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

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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