Real-time fluorescent DNA amplification in a flexible liquid format. Recommended for users who want to combine RPA with the use of TwistDx’s proprietary fluorescent TwistAmp® exo Probes. In addition to the basic components, a powerful nuclease (Exonuclease III) is provided which will process TwistAmp® exo Probes during the amplification reaction itself and generate a real-time readout. The user need only supply primers, probe, dNTPs and template.See manual for more information. Click to order oligonucleotides.
Detail
Real-time fluorescent DNA amplification in a flexible liquid format. Recommended for users who want to combine RPA with the use of TwistDx’s proprietary fluorescent TwistAmp® exo Probes. In addition to the basic components, a powerful nuclease (Exonuclease III) is provided which will process TwistAmp® exo Probes during the amplification reaction itself and generate a real-time readout. The user need only supply primers, probe, dNTPs and template.See manual for more information. Click to order oligonucleotides.
Perfect for: Real-time fluorescent DNA detection, facilitating use of different RPA reaction volumes, or variation of component ratios.
Product code: TALQEXO01
Other Products
cfDNA Purification Kit (Magnetic Beads)
Product Info
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Product Info
cfDNA Purification Kit (Magnetic Beads)
The cfDNA Purification Kit (Magnetic Beads) was developed for cell free DNA (cfDNA) enrichment by separating genomic DNA contamination from isolated cfDNA samples.
Many diagnostic technologies for detection of disease signals in cfDNA begin with isolation and purification of DNA from liquid biopsy that include urine, plasma, cerebrospinal fluid. The most widely explored biotechnology is assays used to detect cancer-derived plasma cfDNA. Silica-based magnetic bead cfDNA isolation kits can reliably extract total DNA from plasma, but typically yield a large variation in cfDNA that includes the presence of genomic DNA that often depends on tumor stage, tumor size, or healthy status individuals. Most of the commercial cfDNA isolation kits can’t specifically recover the cfDNA while leaving the high molecular weight genomic DNA behind. The presence of genomic DNA can lead to decreased sensitivity or inconsistent results in downstream applications such as next-generation sequencing (NGS), PCR, QPCR, and digital PCR etc.
Therefore, an additional purification step to enrich cfDNA before downstream methods helps to improve signal from fragments that originate from cancer cells. A proportion of cancer-derived cfDNA fragment signals are below 100 bp and are often not detectable except by qPCR or single-stranded DNA based library preparation for NGS (1, 2, 3). Furthermore only 1% of cancer-derived fragments are found above 400 bp (1, 2). Capture of size-selected fragments between 90-150 bp improved detection of cancer by 2-4 fold (4). Furthermore, TF-bound and protected cfDNA fragments are also being investigated for active cancer-specific signals down to 35-80 bp (5, 6).
This kit uses Dual Solid Phase Reversible Immobilization (SPRI) technology for cfDNA purification. Most Dual SPRI procedures do NOT recover fragments below 100 bp. The kit can be used for the enrichment of cfDNA isolated from liquid biopsies, plasma, serum, and urine. The kit separates cfDNA (50-500 bp) and genomic DNA, and recovers of 90% of the cfDNA without the high molecular weight genomic DNA with high efficiency. Fragments at 500 bp and above may also be retained. Both the 50-500 bp and >500 bp DNA fractions can be used for downstream applications such as single-stranded or double stranded NGS library prep, qPCR, ddPCR, and other methods.
Features
Separation of cfDNA and genomic DNA; Recovery of both types of DNA
Recovery of cfDNA (50-500 bp)
As short as 50 bp can be recovered
Recovery of high molecular weight genomic DNA
Removal of unwanted components and other impurities
Automation friendly
Examples of cfDNA purification. Both cfDNA and genomic DNA can be recovered separately.
The range of recovered small DNA fragments is from 50 to 500 bp. The input DNA are mixtures of sheared small DNA fragments and intact genomic DNA. The ratios of sheared DNA fragments versus genomic DNA are indicated.
Recovery rates of cfDNA and genomic DNA.
Document
Many diagnostic technologies for detection of disease signals in cfDNA begin with isolation and purification of DNA from liquid biopsy that include urine, plasma, cerebrospinal fluid. The most widely explored biotechnology is assays used to detect cancer-derived plasma cfDNA. Silica-based magnetic bead cfDNA isolation kits can reliably extract total DNA from plasma, but typically yield a large variation in cfDNA that includes the presence of genomic DNA that often depends on tumor stage, tumor size, or healthy status individuals. Most of the commercial cfDNA isolation kits can’t specifically recover the cfDNA while leaving the high molecular weight genomic DNA behind. The presence of genomic DNA can lead to decreased sensitivity or inconsistent results in downstream applications such as next-generation sequencing (NGS), PCR, QPCR, and digital PCR etc.
Fasciolosis is one of the most frequently encountered autochthonous helminthic infections in Central Europe. The immunodiagnosis of Fasciola hepatica infections is challenged by high serological cross-reacitivity with other (hepatic) parasitological infections encountered in Central Europe, such as alveolar echinococcosis, toxocarosis and ascariosis, but also other parasitic infections acquired during overseas travel. The SAP-2 recombinant antigen shows a high specificity, especially with patients with other parasitic infections. It is a suitable serological test for routine diagnosis of human fasciolosis, particularly if the results are supported by clinical history.
This product is manufactured by Bordier Affinity Products in Switzerland and distributed in Germany exclusively by Milenia Biotec.
Method/Platform
ELISA in microplate format
Range/Assay Sensivity
77% sensitivity, 98% specificity
Test Principle
The kit provides all the material needed to perform 96 enzyme-linked immunosorbant assays (ELISA) on breakable microtitration wells sensitized with Fasciola hepaticarecombinant antigen. Specific antibodies in the sample will bind to the antigen and washing will remove unspecific antibodies. The presence of parasite specific antibodies is detected with a Protein A – alkaline phosphatase conjugate. A second washing step will remove unbound conjugate. Revealing bound antibodies is made by the addition of pNPP substrate which turns yellow in the presence of alkaline phosphatase. Color intensity is proportional to the amount of Fasciola hepatica specific antibodies in the sample. Potassium phosphate is added to stop the reaction. Absorbance at 405 nm is read using an ELISA microplate reader.The test can be performed with automatic systems, but this must be validated by the user.
12 x 8 strips (96 tests) Fasciolosis is one of the most frequently encountered autochthonous helminthic infections in Central Europe. The immunodiagnosis of Fasciola hepatica infections is challenged by high serological cross-reacitivity with other (hepatic) parasitological infections encountered in Central Europe, such as alveolar echinococcosis, toxocarosis and ascariosis, but also other parasitic infections acquired during overseas travel. The SAP-2 recombinant antigen shows a high specificity, especially with patients with other parasitic infections. It is a suitable serological test for routine diagnosis of human fasciolosis, particularly if the results are supported by clinical history.
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
Features
5’→3’ DNA polymerase activity
No detectable 3’→5′ exonuclease (proofreading) activity
Generates PCR products with 3′-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
For direct loading into electrophoresis analysis
DNA bands can be visualized directly under UV or 470 nm blue light illumination
Applications
Routine PCR
Colony PCR
High throughput PCR
Amplification of DNA fragments up to 8 kb
Generation of PCR products for TA cloning
DNA labeling
Storage
Protected from light and avoid multiple freeze/thaw cycles 4°C for 6 months -20°C for 24 months
Document
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
