PACE Genotyping Master Mix uses a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals corresponding to genotypes. PACE compatible genotyping assays are comprised of two competitive allele specific forward primers (which differ in their terminal 3’ bases and unique 5’ tail sequences) and a common, reverse primer. PACE Genotyping Master Mix is supplied at 2x concentration and with ROX normalising dye at a range of levels to ensure compatibility with your qPCR instrument or reader.
Genotyping assay designs are available from 3CR Bioscience through our free PACE assay-design service; once designed, users can purchase assay primers independently or through 3CR Bioscience using our partial or full-assay validation service. PACE Genotyping Master Mix is also compatible with KASP™ and Amplifluor® marker assays.
REQUIRED COMPONENTS
qPCR machine or Thermocycler + Fluorescent plate reader
PCR plate or equivalent and appropriate optically clear seal
Template DNA
PCR-grade water
Genotyping assays
For Research and Development purposes only. Not for diagnostic use.
Legal Information KASP™ is a trademark of LGC Biosearch Technologies Amplifluor® is a registered trademark of Merck KGaA
Other Products
Cat.# 20102S, 20102L: Size range 100-200 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
LyoBeads are ready-to-use, freeze-dried master mixes in shape of a small ball or spheres.
Standard LyoBeads contain DNA polymerase(s), reaction buffer and dNTPs. LyoBeads can also contain Primer & Probes when produced customized. They are rehydrated within seconds in any aqueous solutions, which makes reaction setup very simple. Only a biological sample has to be added.
Storage: LyoBeads are shipped and stored simply at room-temperature. This provides a more cost efficient and ecological distribution compared to other master mixes.
LyoBeads can be pre-dispensed in PCR-strips, PCR-plates, cartridges etc.
All necessary components for PCR are already included in one LyoBead: an engineered DNA polymerase, an optimized reaction buffer and ultrapure dNTPs. Only primers and probes need to be added. A hot-start formulation of the included DNA polymerase prevents false amplification during the reaction set-up.
For research use and further manufacturing.
In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
Document
LyoBeads are ready-to-use, freeze-dried master mixes in shape of a small ball or spheres.
Standard LyoBeads contain DNA polymerase(s), reaction buffer and dNTPs. LyoBeads can also contain Primer & Probes when produced customized. They are rehydrated within seconds in any aqueous solutions, which makes reaction setup very simple. Only a biological sample has to be added.
Isolate genomic DNA from animal tissues, cells, bodily fluids, viruses and swabs
Rapid and convenient spin column procedure
Purified DNA is of the highest quality and integrity for sensitive downstream applications including PCR, qPCR, genotyping, sequencing and more
This kit is designed for the rapid preparation of genomic DNA from various tissue samples, cultured cells, viruses, bodily fluids and swabs using a rapid spin column protocol. Purified DNA is of an excellent yield and quality, and is immediately ready for any downstream application including PCR, qPCR, genotyping, sequencing and more. The protocol can be completed in approximately 80 minutes (including incubation time).
Average Yield:* HeLa Cells (1 x 106) Tissue (from 10 mg kidney)
8 µg 10 µg
Maximum Amount of Starting Material: Animal Tissues Cultured Cells Bodily Fluids (blood, saliva) Viral Suspension
20 mg 3 x 106 cells 150 µL 150 µL
Time to Complete 10 Purifications
80 minutes
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
