Introduction

The HiPure Plant RNA Midi Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 1mg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from <1g simple plant sample without chloroform
Applications	RT-PCR, qPCR, Northern hybridization, second generation sequencing, nucleic acid protection, in vitro translation
Purification method	Midi spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	Plant leaves, roots, stems, fruits, fungi, etc.
Sample amount	0.5-1 g
Elution volume	≥400μl
Time per run	≤40 minutes
Liquid carrying volume per column	4ml
Binding yield of column	1mg

Principle

The Kit isolates total RNA from up to 1g plant tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 40 min. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 400µl water using the Kit.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – medium isolation of several samples can be completed in 40 minutes
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• Broad spectrum – various types of plant samples can be processed by diversity of operating procedures

Kit Contents

Contents	R415202	D415203
Purification Times	20 Preps	100 Preps
HiPure DNA Midi Column II	20	100
HiPure RNA Midi Columns II	20	100
15ml Collection Tubes	40	200
Buffer RLC	120 ml	550 ml
Buffer PRC1	120 ml	550 ml
Buffer RW1	70 ml	400 ml
Buffer RW2*	20 ml	100 ml
RNase Free Water	30 ml	120 ml

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. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the Buffer RLC/PRC1. Dissolve by warming buffer to 37°C.

Purchase Guide

1. When dealing with woody or uncommon samples, R4150 is recommended first. R4150 contains two polysaccharide/polyphenol lysis buffer, which is the most universal product.

2. R4151 is recommended for handling common economic crop samples for the first time. Strong lysis solution can be used to process easy-extraction samples. The amount of corn or rice leaves samples can reach up to 300mg.

3. R4165 adopts CTAB/chloroform method, which can also handle a large number of difficult-to-extraction plants, but requires contact with chloroform substitutes, which is less safe than other kits. This kit uses DNase Ⅰ to remove DNA, which is also a good choice for extracting polysaccharide/polyphenol-rich plant samples.

4. R4014 is recommended for fruit/starch plant samples, which uses improved trizol pre-treatment, single column operation and is more economical.

Select the right purification kit to get impactful results：

Ⅰ. Purchase guide of Magen kits based on sample

Ⅱ. Cross purchase guide for Magen kits vs Other brands

For any customized product or OEM service, please contact us.

The HiPure Plant RNA Midi Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 1mg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Overview

• ASTM Type I Water
• Produced by reverse osmosis, ion-exchange, electro-deionization, ultrafiltration, UV treatment (UVC & 185 nm), 0.1 μm membrane filtration
• Nuclease-free, protease-free, endotoxin-free
• Suitable for molecular biology applications (qPCR, NGS etc.)
• Suitable for cell culture applications
• Produced in an ISO certified GMP laboratory
• Water system is routinely monitored for compliance with QC standards

The Norgen water purification system has been designed in conjunction with experts from Millipore Sigma. The Ultrapure NFW is purified in a two-stage process. The first stage, to produce type II water, includes pre-treatment with activated charcoal and polyphosphate, this water then undergoes reverse osmosis, electro-deionization and ion exchange to remove contaminants as well as UV treatment for disinfection. The Pure water then flows to a second system in a clean room with an additional 4 layers of purification. After passing through an additional ion exchange column, the water is then subjected to Ultrafiltration to remove particulates, bacteria, & ionic contaminants down to trace level, the water is then treated with UV light (182 nm wavelength) to break down organic matter. Lastly a second Ultrafilter for the production of pyrogen-, nuclease- and bacteria-free water at 18.2 MΩ. This water is of utmost purity and suitable for direct use in molecular biology applications such as qPCR and NGS, as well as cell culture and other applications.

Product Specifications
pH	n/a (highly pure water does not contain enough ions for an exact pH determination.)
Recommended Storage	Room Temperature
Grade	Ultrapure (ASTM Type I)
Biological Activity	DNase-, RNase-, Protease-, Endotoxin- Free
TOCs	< 50 ppb
Resistivity	> 18 MΩ-cm
Treatment	Not DEPC-Treated
Quantity	100 mL, 500 mL, 1000 mL
Purification Methods	Activated charcoal, polyphosphate, reverse osmosis, electro-deionization, ion exchange, UV, ultrafiltration
Shipping Conditions	Room Temperature

Details

Applications
For use in any molecular biology application

Storage Conditions
Norgen’s Nuclease-Free Water should be stored at room temperature. Storage at +4°C or -20°C is optional.

Precautions and Disclaimers
This product is designed for research purposes only.  It is not intended for human or diagnostic use.

OverView

HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human  total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results  indicate that HL-dsDNase has minimal impact on RNA quality and quantity.

HL-dsDNase is an engineered version of dsDNase that is rapidly and completely inactivated by incubation for 5 minutes at 58°C with 1 mM DTT at pH 8.0 or above. Chemical inactivation in downstream compatible RT or PCR buffers allows milder heat inactivation and, in some cases, skipping heat inactivation altogether. This makes HL-dsDNase very useful for removal of DNA from RNA preparations since the enzyme may be inactivated using various strategies with reduced risk of auto-degradation of  RNA in the presence of magnesium, making HL-dsDNase an ideal enzyme when working with small volumens of RNA.

HL-dsDNase is also an excellent choice for removal of unwanted external DNA from samples prior to analysis of nucleic acids protected by biological membranes, e.g., bacteria, viruses, and sperm. Especially if using downstream analysis methods that might be affected by host cell DNA, as metagenomic sequencing and STR-profiling. The easy inactivation of HL-dsDNase makes it a fast and efficient alternative to methods as differential extraction, where sample material is often lost.

Recommended applications:

• Removal of genomic DNA from RNA preparations
• Improved removal of host-cell DNA from sperm cells prior to STR-profiling

Key advantages with HL-dsDNase:

• Double-strand DNA specific endonuclease
• Easily heat-inactivated by very moderate heat treatment
• High specific activity
• Useful for removal of DNA from RNA preps

Properties

HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human  total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results  indicate that HL-dsDNase has minimal impact on RNA quality and quantity.