Solid Phase Reversible Immobilization (SPRI) beads are a simple and effective reagent for DNA purification, but not for short oligo purification. The beads are paramagnetic particles coated with carboxyl groups that can reversibly bind to nucleic acid. However, SPRI beads can only purify DNA/RNA fragments that are 100 base pairs or longer. DNA/RNA fragments shorter than 100 base pairs are not effectively recovered.
Detail
Magnetic Beads (Short Oligo Purification)
Solid Phase Reversible Immobilization (SPRI) beads are a simple and effective reagent for DNA purification, but not for short oligo purification. The beads are paramagnetic particles coated with carboxyl groups that can reversibly bind to nucleic acid. However, SPRI beads can only purify DNA/RNA fragments that are 100 base pairs or longer. DNA/RNA fragments shorter than 100 base pairs are not effectively recovered.
Oligo purification can also be performed using spin column-based technology. The oligo size limitation for recovery is around 20 nt, as oligos under 20 nt have a very low recovery rate.
We have developed Magnetic Beads (Short Oligo Purification) Kit for short oligo purification. Our proprietary bead technology enables the recovery of oligos as short as 6 nt. 80% of the 6 nt oligos and 90% of the > 8 nt oligos can be recovered. The reagent also effectively removes impurities and unwanted components such as salts, proteins, dNTPs, detergents, and other contaminants. The magnetic bead reagents are RNase free, and can be used for both DNA and RNA applications.
Recovery rates of of short oligos. Oligos with different sizes were used as input. BioDynami Short Oligo Quantification Kit (Cat. # 40046) was used to quantify the recovery rates with modifications.
Features
Effective purification of short oligos
6 nt oligos: 80% recovery rate
>8 nt oligos: >90% recovery rate
Removal of impurities and unwanted reaction components
Other Products
Cat.# 20101S, 20101L: Size range 50-100 bp
Product Info
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Product Info
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
Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. The extracted products can be used for clinical in vitro detection.
Details
Specifications
Features
Specifications
Main Functions
Isolation circulating DNA from 1-5ml plasma, serum, body fluids using vacuum protocol
Applications
qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (vacuum)
Sample type
Serum, plasma and other cell-free fluid samples
Sample amount
1-5ml
Elution volume
≥50μl
Time per run
≤60 minutes
Liquid carrying volume per column
4ml
Binding yield of column
1mg
Principle
This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer.
Advantages
High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
High concentration – low elution volume, ensuring high nucleic acid concentration
High purity – low alcohol binding method, completely removing inhibitor and protein pollution
High recovery – DNA can be recovered at thelevel of PG by silica gel column purification
Kit Contents
Contents
IVD3182
Purification Times
50
Buffer ACL
250 ml
Buffer ACB*
300 ml
Buffer DCW1*
22 ml
Buffer DCW2*
10 ml
Proteinase K
540 mg
Protease Dissolve Buffer
30 ml
Carrier RNA
110 μg
Nuclease Free Water
20 ml
HiPure CFDNA Mini Columns
50
2 ml Collection Tubes
100
Extender Tube
50
Vac-Connector
50
Storage and Stability
Proteinase K, Carrier RNA 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 dry 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
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Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,
Lateral Flow Assay designed for the detection of IgG antibodies against PF4/polyanion-complexes in human citrated plasma or Serum
This product is only available in Germany!
Method/Platform
lateral flow, immunoassay
Range/Assay Sensivity
Test Principle
IgG antibodies are resposible for the heparin induced thrombocytopenia.
Immobilized anti-human IgG on the membrane of the test unit binds patients IgG-antibodies which are previously captured by the PF4/polyanion-complex which is detected by intensely colored gold nanoparticles.
The presence of PF4/polyanion-complex becomes visible at a colored test line. The surplus of gold particles continues to migrate through the membrane and is captured at the control line by specific antibodies.
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Lateral Flow Assay designed for the detection of IgG antibodies against PF4/polyanion-complexes in human citrated plasma or Serum
