Introduction

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.

Details

Specifications

Principle

The Kits are designed to purify both genomic DNA and total RNA from the same cellor tissue sample. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column and bind DNA. Ethanol is added to the flow-through and the sample is applied to an RNA column. DNA/RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit. High-quality DNA is eluted in as little as 50µl water using the Kit.

Advantages

• High quality – high purity total RNA / DNA can be directly used in a variety of downstream applications
• Fast – column method can complete the extraction of several samples in 30 minutes
• Safe – no phenol chloroform extraction required
• Simultaneous extraction- simultaneously isolate DNA and RNA from one sample

Kit Contents

Storage and Stability

HiPure DNA/RNA Kit can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

Magnetic Beads (DNA & RNA Purification)

Solid Phase Reversible Immobilization magnetic beads consist of paramagnetic particles coated with carboxyl groups that reversibly bind DNA. They are used for DNA purification because they are fast, simple and efficient. We have developed our own beads technology that are different from other SPRI beads technologies.

Our Magnetic Beads (DNA & RNA Purification) combines BioDynami’s proprietary chemistries with reversible DNA-binding properties of magnetic beads. The magnetic beads, which are unique from other SPRI beads, are developed for effective nucleic acid purification by removing unwanted components such as salts, dNTPs, enzymes, primers, adapters, and other impurities. The beads are RNase free, can be used for applications of DNA, and even work with more sensitive RNA without any additional cost.

Our magnetic beads are optimized to selectively bind DNA fragments of 100 bp and larger, and RNA fragments of 200 bases and larger, similar to other SPRI beads such as AMPure® XP* and SPRIselect*. Purified DNA and RNA are suitable for downstream applications requiring high quality DNA and RNA, as the purified fragments are free of contaminants and impurities. The beads can be used for NGS library purification, PCR fragment cleanup, molecular cloning, or even nucleic acid concentration.

The beads can also be used for size selection of DNA fragments ranging from 150 bp to 800 bp by changing the bead-to-sample volume ratio and performing single or double-size selection. The beads are an ideal choice for NGS library preparation. They can be easily integrated into the standard workflow of NGS library preparation since the volume ratio is similar for protocols using standard magnetic beads.

Features:

• Effective recovery of DNA and RNA samples
  • DNA fragments greater than 100 base pairs
  • RNA fragments greater than 200 bases
• Removal of unwanted components and impurities
• Fragment size selection for specific applications
  • Consistent single or double-size selection
• Flexibility: compatible with manual and automated processing
• Cost effective alternative to other beads such as AMPure® XP* and SPRIselect*  with equivalent performance

* AMPure® XP and SPRIselect are trade marks of Beckman Coulter.

Magnetic beads recovery rate. dsDNA and ssDNA of genomic DNA, 1 kb DNA, and 200 bp DNA fragments were used. Total RNA was also tested.

Comparison of elution volume vs yield. 40 ul , 30 ul, and 20 ul of elution volume were used. BioDynami magnetic beads have better recovery rates at low elution volume when compared to other SPRI beads such as AMPure® XP*.

Applications:

• NGS Library preparation
• Purification of PCR fragments
• Purification of DNA and RNA fragments
• Molecular Cloning
• Other applications requiring purified DNA and RNA

The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.

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

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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. 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 technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.

Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.

DNA size selection with dual clean-ups.

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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.

DNA size selection with single clean-up for >5 kb and >10 kb DNA.

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Features of DNA size selection and library size selection

• • • High specificity and high recovery of size selection
    • 11 selection ranges are available, including 5 ranges for NGS library size selection
      • • 50-100 bp
        • 100-200 bp
        • 200-500 bp
        • 250-350 bp: ideal for illumina PE100 sequencing
        • 300-450 bp: ideal for illumina PE150 sequencing
        • 450-750 bp: ideal for illumina PE300 sequencing
        • 500-1000 bp
        • 1-3 kb
        • 1-5 kb
        • >5 kb: ideal for long-read sequencing
        • >10 kb: ideal for long-read sequencing
    • Fast and simple
      • • 20-min protocol
        • No gel purification required
        • No columns required
        • No centrifugation required
    • Efficient removal of contaminants and unwanted components