Introduction

HiPure FFPE Nucleic acid Kit supplies a simple and rapid DNA extraction for formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 20 minutes (not including digestion time). DNA can be directly used for downstream applications such as PCR, southern blot and viral DNA detection, etc.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from FFPE tissue samples
Applications	PCR, southern blot and viral DNA detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples
Sample amount	<20mg
Elution volume	>15µl
Time per run	≤20 minutes
Liquid carrying volume per column	4ml
Binding yield of column	100μg

Principle

Hipure FFPE Nuclear acid kit adopts silica gel column purification. The sample is deparaffinated by xylene and digested by lysate and protease. After de crosslinked at 90 ℃, DNA/RNA is released into the lysate. Adding ethanol to adjust the binding conditions, the sample is transferred to the column where DNA/RNA is adsorbed on the membrane and protein is removed without adsorption. Protein and other impurities are washed by buffer GW1, and the salt is removed by buffer GW2. Finally, the DNA / RNA is eluted by low salt buffer.

Advantages

• Safety – deparaffinating without contacting with xylene or other toxic solution
• Fast – without overnight incubation and digestion, several samples can be extracted within 2hours
• High efficiency -remove the formaldehyde modification of DNA, greatly enhancing the sensitivity of PCR

Kit Contents

Contents	D312602	D312603
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns I	50	250
2ml Collection Tubes	50	250
Buffer DPS	30 ml	150 ml
Buffer ATL	15 ml	60 ml
Buffer AL	15 ml	60 ml
Buffer GW1*	22 ml	88 ml
Buffer GW2*	12 ml	50 ml
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	10 ml	30 ml

Storage and Stability

Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. 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.

Experiment Data

HiPure FFPE Nucleic acid Kit supplies a simple and rapid DNA extraction for formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 20 minutes (not including digestion time). DNA can be directly used for downstream applications such as PCR, southern blot and viral DNA detection, etc.

DBCO-PEG6-Maleimide is a PEG linker containing a DBCO moiety and a terminal primary maleimide group. The maleimide group will react with a thiol group to form a covalent bond, enabling the connection of biomolecule with a thiol. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG8-Maleimide is a PEG linker containing a DBCO moiety and a terminal primary maleimide group. The maleimide group will react with a thiol group to form a covalent bond, enabling the connection of biomolecule with a thiol. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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.

.

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.

.

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.

.

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