NGS Library Quantification Standards With PCR Primers (Ion Torrent Platform)
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The NGS Library Quantification Standards with PCR Primers (for Ion Torrent platform) were developed for quantifying the library concentration for ion torrent sequencing platform. Quantification of the library of the fully ligated libraries is important for the quality of the sequencing outcome. Optimal library concentration can increase sequencing yield. Poor library concentration results in bad emPCR, which can lead to low sequencing capacity.
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The NGS Library Quantification Standards with PCR Primers (for Ion Torrent platform) were developed for quantifying the library concentration for ion torrent sequencing platform. Quantification of the library of the fully ligated libraries is important for the quality of the sequencing outcome. Optimal library concentration can increase sequencing yield. Poor library concentration results in bad emPCR, which can lead to low sequencing capacity.
QPCR is the best method for library quantification. Our reagent only amplifies library molecules that will be used for subsequent emPCR, and is optimized for amplification of various samples. Our reagent is compatible with commercial SYBR Green based QPCR reagents. Quantification of library concentration is achieved by comparison with a standard curve generated from DNA Standards.
The kit comprises DNA Standards (six 10-fold dilutions) and a primer mix.
NGS Library Quantification Standards with PCR Primers (Ion Torrent platform): real time quantitative PCR curve of the standards.
Attogene’ s RNase Inhibitor is a protein-based ribonuclease inhibitor which noncovalently binds and inactivates a wide variety of RNases in a range of temperature (37–65°C) and pH (5.5–8.5) conditions. This product is distinct from placental RNase Inhibitor protein in that it inactivates RNases I and T1 in addition to RNases A, B and C. NA2021 is distinct from RNase Inhibitor protein in that it is less expensive, have more robust interactions with RNAses.
MADE AND FUNCTIONALLY TESTED AT ATTOGENE (MADE IN AUSTIN TEXAS – USA)
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RNase Inhibitor (10,000 units)
Concentration: 40 units/μL
Volume: 0.25 mL
Storage Conditions: Store at 2°C – 8°C
Inhibits: RNases A, B, and C
Supplied in: 1X PBS with 0.05% Sodium Azide.
Brucella abortus is an intracellular, blood-borne parasite. It is a Gram-negative coccobacillus that causes an infectious and contagious disease called Brucellosis. The disease primarily affects cattle but it can also be transmitted to humans from infected animals and consuming their products. The disease can lead to great economic loss especially in the dairy and agricultural industry. The Brucella abortus genome contains two DNA chromosomes in a circular confirmation; the first chromosome is approximately 2.1 Mb and the second chromosome is approximately 1.2Mb. Unusually it does not contain any plasmids or genomic islands that relate to pathogenicity and lacks many other genes that code for common virulence factors including capsules, fimbriae, exotoxins, cytolysins, resistance forms, or antigenic variation. The most common mode of transmission to humans is through the ingestion of unpasteurized milk and cheese products as the bacteria are present in the milk glands of infected female cows. In cattle transmission can also be through ingestion but in addition, the bacteria can persist in the reproductive tracts of males, namely seminal vesicles, ampullae, testicles, and epididymides, allowing sexual transmission. In humans the bacteria enter macrophages by phagocytosis and then live in compartments of vacuolar space along the endoplasmic reticulum. They persist by inhibiting host apoptosis and go onto form chronic disease causing lesions in the liver, spleen, bone marrow and kidneys. In cattle the bacteria additionally infect the trophoblast epithelial cells, which provide nutrition to the embryo. The trophoblast cells eventually lyse, releasing further bacteria into the blood stream of the embryo. The B. abortus cells in the blood stream go on to colonize the placenta and fetus in pregnant female cows, resulting in abortion of the fetus. Abortion can also result from insufficient anti-Brucella activity in the amniotic fluid. In humans, the disease can be either acute or chronic and some of the symptoms include fluctuating fever, chills, sweating, headache, muscle pain and weight loss. Once a person becomes infected they are prescribed a combination of tetracycline and streptomycin for 3-6 weeks. In cattle, additional symptoms include arthritic joints and retained after-birth.
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Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target
Accurate controls to confirm findings
The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
PCR-Free NGS DNA Library Prep Kit Workflow
PCR amplification is a standard step for library preparation of Next-Generation Sequencing. The PCR is used to amplify fully ligated DNA fragments and to add index information to the libraries. The indexing is for the pooling of the library samples in an effort to reduce the sequencing cost.
However, PCR introduces uneven amplification of DNA in some DNA regions with extreme GC-contents and secondary structures. This bias can cause very low sequencing coverage in such regions. DNA sequencing in these regions is still a huge challenge.
PCR-free library prep can reduce library bias and minimize sequencing gaps. The sequencing data from PCR-free library samples have even genomic coverage with few gaps and better depth in GC-rich regions. Our PCR-free NGS kit offers optimal coverage in the regions that are traditionally difficult, such as high-GC content regions, low-GC content regions, and repetitive sequence regions.
Two index types are available for the kit:
Non-index: Libraries do not have index.
Index: Each library contains one i5 index and one i7 index. Library multiplexing up to 96 samples is possible. List of indexes can be downloaded Here.
Kit advantages:
Total time: 1 hr
Hands-on time: 5 min
Easy procedure: Ready-to-use master mix & Less reaction components
Input DNA amount from 100 ng to 1 ug
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The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
