Introduction

This product provides fast and easy method for purification of high purity DNA from bone samples. The obtained DNA can be directly used for PCR and STR detection down stream application.

Details

Kit Contents

Contents	D312402	D312403
Purification Times	50	250
HiPure DNA Mini Columns I	50	250
2ml Collection Tubes	50	250
Buffer BGL	40 ml	180 ml
Buffer GXP	30 ml	150 ml
Buffer GW1*	13 ml	53 ml
Buffer GW2*	20 ml	2 x 50 ml
DTT Powder	235 mg	2 x 235 mg
Proteinase K	48 mg	240 mg
Protease Dissolve Buffer	5 ml	15 ml
Elution Buffer	15 ml	60 ml

Storage and Stability

This product can be stored at room temperature (15~25°C) for 18 months. Proteinase K/DTT dry powder can be transported and stored at room temperature. For long-term storage (>6 months), it is recommended to store at -20~8°C. Dissolved Proteinase K should be stored at -20~8°C. The dissolved DTT should be stored at -20°C.

This product provides fast and easy method for purification of high purity DNA from bone samples. The obtained DNA can be directly used for PCR and STR detection down stream application.

Overview

• Purify amplified DNA ranging from 100 bp -15,000 bp in size
• Fast and efficient spin column format 
• Available in a 50 prep size and a 250 prep size
• Also available in 96 well format
   

This kit enables the rapid purification of amplified DNA products from PCR mixes. It is able to effectively remove PCR by-products including primers, dimers, enzymes, unincorporated nucleotides and mineral oil from the desired PCR product. The purified PCR products are fully compatible with restriction enzyme digestion, ligation into vectors, labeling, sequencing and more. This kit can also be used as an alternative to organic extraction and ethanol precipitation to clean up various enzymatic reactions.

The kit is also available in a 96-well format for high-throughput PCR purification. Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.

Details

Supporting Data

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Kit Specifications – 96-well
Binding Capacity Per Well	15 μg
Maximum Loading Volume Per Well	500 μL
Size of DNA Purified	100 – 15,000 bp
Average DNA Recovery	> 90%
Average Primer Reoval	> 90%
Minimum Elution Volume	50 μL
Time to Complete 96 Purifications	20 minutes

 
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 14400 (50 preps)	Cat. 45700 (250 preps)	Cat. 24800 (192 preps)
Binding Buffer C	30 mL	5 x 30 mL	3 x 30 mL
Wash Solution A	12 mL	2 x 20 mL	2 x 38 mL
Elution Buffer B	8 mL	2 x 30 mL	2 x 15 mL
Spin Columns	50	250	–
Collection Tubes	50	250	–
96-Well Plate	–	–	2
Adhesive Tape	–	–	4
96-Well Collection Plate	–	–	2
Elution Tubes (1.7 mL)	50	250	–
96-Well Elution Plate	–	–	2
Product Insert	1	1	2

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