Introduction

Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:

1. Humic acid pollution. The soil, especially in forests and grasslands, is rich in humic acids. Humic acid is a series of organic molecules, some of which are very similar to nucleic acid molecules and difficult to remove during purification. Trace amounts of humic acid pollution can lead to downstream applications such as PCR and enzyme digestion failure.

2. Lysis method. Soil samples contain various microorganisms, such as bacteria and fungi. Gram positive bacteria and fungi both contain very thick bacterial walls, and effectively breaking down the cell walls of these microorganisms is crucial for extracting high-yield metagenomic DNA. Due to the complexity of soil samples, it is not feasible to use enzymatic methods (such as lysozyme, wall breaking enzyme, snail enzyme) or liquid nitrogen grinding, as the soil contains various metalions or inhibitory factors that inactive the digestive enzymes, or the presence of sand particles in the soil makes liquid nitrogen grinding difficult.

3. The DNA yield is difficult to control. Soil samples would have significant changes in the number and variety of microorganisms due to fertility, inferiority, high moisture content, dryness, or depth of sampling. In a small range of soil samples, the DNA content often varies by thousands of times. In addition, certain chemical components in soil, such as heavy metal salts and clay substances, can cause a decrease in DNA yield.

Magen’s HiPure Soil DNA Kits are currently the most optimized kit for soil DNA extraction. The kit adopts glass bead grinding method and thermal shock chemical wall breaking method, which can be carried out in the point vortex instrument without special bead grinding instrument, and is suitable for a wide range of laboratories. The Absorber Solution in the reagent kit is a humic acid adsorbent exclusively developed by Magen Company, which can efficiently remove various humic acid pollutants. In addition, an alcohol-free silica gel column purification method is also used to efficiently remove various soluble metal salts and other soluble inhibitory factors from the soil. The kit has successfully extracted from the following soil (partially based on customer feedback): soil from forests in nature reserves (30 to 40 years old forest soil with a surface layer of 30-50cm deciduous layer), mangrove soil, grasslands, farmland, seabed mud, sludge, mineral area soil, organic matter contaminated soil, pond mud, garbage mud, air conditioning pipeline deposits, etc.

This product allows rapid and reliable isolation of high-quality genomic DNA from various soil samples. Up to 500 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatilityof spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Details

Specifications

Features	Specifications
Main Functions	Isolation DNA from 200-500mg soil sample
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Soil
Sample amount	200-500mg
Elution volume	≥30μl
Time per run	≤60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

Soil sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. humic acid,proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Advantages

• Fast – several samples can be extracted in 40 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• Good repeatability – silica technology can obtain ideal results every time
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D314202	D314203
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns II	50	250
2ml Collection Tubes	50	250
2ml Bead Tubes	50	250
Buffer SOL	60 ml	250 ml
Buffer SDS	5 ml	20 ml
Buffer PS	10 ml	50 ml
Absorber Solution	10 ml	50 ml
Buffer GWP	40 ml	220  ml
Buffer DW1	30 ml	150  ml
Buffer GW2*	20 ml	2 x 50  ml
Buffer AE	15 ml	30 ml

Storage and Stability

Absorber Solution 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.

Experiment Data

Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:

200 µL filtered pipette tips for the Opentrons Flex, provided sterile in racks or blister packs (refills). Tips are clear, non-conducting polypropylene tips, free of human DNA, PCR inhibitors, RNase/DNase, pyrogens, and endotoxins. Opentrons Flex Tips and Racks are autoclavable.

200 µL filtered pipette tips for the Opentrons Flex, provided sterile in racks or blister packs (refills). Tips are clear, non-conducting polypropylene tips, free of human DNA, PCR inhibitors, RNase/DNase, pyrogens, and endotoxins. Opentrons Flex Tips and Racks are autoclavable.

Introduction

The HiPure Fruit RNA Mini Kit provides fast purification of high-quality RNA from fruits, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100µg 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 <150 mg fruit or seed using chloroform and Plantzol reagent
Applications	RT-PCR, qPCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation translation
Purification method	Mini spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	Simple plant, petal, peel, seed
Sample amount	50-150mg
Elution volume	≥300μl
Time per run	≤15 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

The Kit isolates total RNA from up to 150mg plant tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 15 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 an RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 300µl water using the Kit.

Advantages

• Efficient DNA removal – one step RNA extraction can effectively remove genomic DNA
• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 15 minutes by column method
• Wide range – samples including animals, plants, bacteria, cells, etc
• High output – large sample amount, and the yield can be up to 200μg

Kit Contents

Contents	R401402	D401403
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
PlantZol Reagent	60 ml	200 ml
Buffer GXP	30 ml	150 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 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. 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.

The HiPure Fruit RNA Mini Kit provides fast purification of high-quality RNA from fruits, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100µg 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.