Introduction

HiPure HP Plant RNA Mini Kit combines guanidine isothiocyanate lysing and silica gel membrane purification technology to simplify total RNA extraction. Ultracentrifugation in CsCl purification and LiCl / ethanol precipitation are not required. The kit uses DNase digestion to completely remove DNA. It is suitable for extracting up to 100μg of total RNA (including miRNA) from plant samples less than 200mg. Several samples can be extracted within 40 minutes. The purified RNA can be directly used for RT-PCR, fluorescent quantitative RT-PCR, Northern hybridization, second generation sequencing, etc.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA (include miRNA)  from <200mg difficult-to-extract plant samples (use low toxicity chloroform substitutes)
Applications	RT-PCR, qPCR, Northern hybridization, second generation sequencing, nucleic acid protection, in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Hard-to-extraction plant samples such as fruit and seed, grape leaves, tea
Sample amount	≤200 mg
Elution volume	≥30μl
Time per run	1-24 samples within 30 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This kit uses glass fiber filter membrane purification technique, and only requires simple combination-washing-elution steps. The sample is lysed and homogenized in the solution containing guanidine salt, ethanol is added to provide appropriate binding conditions, and transferred to the purification column for centrifugation. Up to 100µg of RNA can be selectively bound to the membrane, pollutants are efficiently washed off after three times of washing, and finally the purified RNA is eluted by RNase Free Water.

Advantages

• Completely remove DNA by using of DNase
• 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 by column method
• 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	R416502	D416503
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	250
DNase I	600 μl	5 x 600 μl
DNase Buffer	6 ml	30 ml
Buffer PAL	60 ml	270 ml
Buffer GXP2*	20 ml	100 ml
Buffer BDP	60 ml	270 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

DNase I should be stored at -20-8°C upon arrival. 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.

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.

HiPure HP Plant RNA Mini Kit combines guanidine isothiocyanate lysing and silica gel membrane purification technology to simplify total RNA extraction. Ultracentrifugation in CsCl purification and LiCl / ethanol precipitation are not required. The kit uses DNase digestion to completely remove DNA. It is suitable for extracting up to 100μg of total RNA (including miRNA) from plant samples less than 200mg. Several samples can be extracted within 40 minutes. The purified RNA can be directly used for RT-PCR, fluorescent quantitative RT-PCR, Northern hybridization, second generation sequencing, etc.

Gel images of different ranges of library size selection. Sheared human genomic DNA was used as input.

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Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.

There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.

Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.

The magnetic beads, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.

Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.

NGS library size selection with dual clean-ups.

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Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.

NGS library size selection with single clean-up for >5 kb and >10 kb libraries.

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Features of NGS library size selection

• • • 5 ranges for NGS library size selection
      • • illumina PE100 sequencing: 250-350 bp
        • illumina PE150 sequencing: 300-450 bp
        • illumina PE100 sequencing: 450-750 bp
        • Long-read sequencing: >5 kb
        • Long-read sequencing: >10 kb
    • Rapid protocol: only 20 minutes
    • Simple workflow
      • • No columns required
        • No gel purification required
        • No centrifugation required

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