Mastitis is the single most costly disease of dairy cattle resulting in the reduction of milk yield and quality. The inflammation of the udder is mainly caused by infection of various bacteria. One of such mastitis bacteria, Streptococcus agalactiae, is highly infectious and causes mainly subclinical infections, which are not identified by the herdsman. As a result, S. agalactiae can spread widely within a herd, causing immediate loss due to reduced milk. S. agalactiae is a gram-positive bacteria belonging to the Group B streptococci. Traditional cultural identification of S. agalactiae is based on S. agalactiae being beta-hemolytic as well as presence of group B Lancefield antigen and by its ability to hydrolyze sodium hippurate.
Storage Conditions and Product Stability All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request
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
ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.
Features
Contains all components for reverse transcription
High yield
Thermostable, up to 50°C, during first strand synthesis
High processivity, generating cDNA up to 8 kb
Reduced RNase H ribonuclease activity
Application
Generation of first strand cDNA from total RNA or mRNA.
Suitable for generating cDNA from RNA with strong secondary structure which can be reduced at temperature up to 50°C.
Storage
-20°C for 24 months
High yield
High sensitivity
Thermostable, up to 50°C, during first strand synthesis
Contents
ComponentVolume ExcelRT™ Reverse Transcriptase (200 U/μl) 100 μl RNase Inhibitor (20 U/μl)100 μl 5X RT Buffer (DTT)500 μl dNTPs Mix (10 mM each)200 μl Oligo (dT)20 (50 μM)100 μl Random Hexamers (100 μM)100 μl DEPC-Treated H2O 1 ml x 2
Storage Buffer
Reverse Transcriptase: 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 1 mM DTT, stabilizer and 50% (v/v) glycerol
RNase Inhibitor: 40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT, 0.1 mM EDTA, stabilizer and 50% (v/v) glycerol
5X RT buffer (DTT)
250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 and 50 mM DTT
Storage
-20°C for 24 months
Document
ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.
