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The Tetro cDNA Synthesis Kit contains our highly sensitive MMLV reverse transcriptase, oligo (dT)18 and random hexamer primers and all the necessary components to generate high quality cDNA from RNA templates (fig. 1). The first-strand cDNA generated is ideal for PCR (fig. 2) and can be used in a variety of other applications, such as analyses of cellular RNAs, characterization of RNA splice variants and the generation and cloning of cDNA. The Tetro cDNA Synthesis Kit is optimized for RT reactions over a wide range of total RNA concentrations (10 pg-2 μg), such that long and low-abundance cDNAs can be detected by amplification after cDNA synthesis. The kit contains oligo (dT)18 and random hexamer primers. The kit components are fully optimized to generate maximum yields of full-length cDNA.
ImmoMix™ is a complete ready-to-use high yield (fig. 1), heat-activated 2x reaction-mix, which simply requires the user to add water, template and primers, and then pre-heat to 95°C for 10 minutes to successfully carry out PCR assays. The 10 minute activation step eliminates the presence of non-specifics such as primer-dimers and mis-primed products, since the enzyme is inactive at initial low temperatures. ImmoMix is based on IMMOLASE™ DNA Polymerase, which leaves an ´A´ overhang, and has been optimized for a wide variety of templates. Additional MgCl2 solution is included should any fine adjustments be required. ImmoMix reduces the time needed to set up reactions, thereby reducing the risk of contamination. Greater reproducibility is ensured by reducing the number of pipetting steps that can lead to pipetting errors.
To complement the SensiFAST™ Probe and SYBR® kits, Bioline has developed the SensiFAST™ cDNA Synthesis Kit to provide a rapid and sensitive method for first-strand cDNA synthesis. The SensiFAST™ cDNA Synthesis Kit displays excellent linearity across a wide range of starting material, revealing the same relative representation in cDNA templates, regardless of gene abundance, making it excellent for use in real-time PCR studies. A novel, highly-pure reverse transcriptase and new TransAmp™ buffer system delivers highly-efficient synthesis of cDNA, enhanced reproducibility and data accuracy. These features make the SensiFAST™ cDNA Synthesis Kit ideal for working with limited samples, such as laser-micro dissected samples and tissue biopsies. The TransAmp Buffer also employs a unique blend of random hexamer primers and anchored oligo dT to ensure unbiased 3’ and 5’ coverage and reverse transcription of all regions. Additionally, the SensiFAST™ cDNA Synthesis Kit can be used with SensiFAST™ Probe and SYBR® Kits for fast real-time RT-PCR without compromising quality, with real-time results in less than an hour. "Our Institute consisting of more than 60 active researchers has a longstanding interest in understanding changes to the transcriptional landscape of prostate cancer. Reproducibility, sensitivity and cost-effectiveness are critical parameters in selecting our day-to-day cDNA synthesis kit. This combined with its convenient two reagent system continues to make the SensiFast cDNA synthesis kit stand out from its competitors and our preference for almost two years." Martin Sadowski, Queensland University of Technology, Brisbane, Australia
Product Highlights Reproducible – consistent results between technical replicates for increased confidence in results Specific – antibody-mediated hot-start DNA polymerase minimizes non-specific amplification for improved assay sensitivity and reliability Sensitive – reliable quantification of low abundance targets and scarce samples Robust – reliable, accurate detection of DNA and RNA targets from a broad range of sample types Fast – delivers reproducible, accurate assay results in as little as 30 minutes Product Description The SensiFAST™ SYBR Lo-ROX Kit has been developed for fast, highly accurate real-time PCR and has been validated on all commonly-used real-time instruments that require a low concentration of the passive reference dye ROX. A combination of the latest advances in buffer chemistry and PCR enhancers ensures that the SensiFAST SYBR Lo-ROX Kit produces reliable assay results under fast thermal cycling conditions. An antibody-mediated hot-start DNA polymerase system promotes highly-specific amplification, in turn improving assay sensitivity and dynamic range. The SensiFAST SYBR® Lo-ROX Kit has been optimized to deliver optimal performance in tandem with the SensiFAST cDNA Synthesis Kit, which offers fast, unbiased cDNA synthesis, without compromising cDNA yield or coverage.
HyperPAGE II is a broad range prestained protein marker containing nine highly purified recombinant proteins with molecular weights ranging from 10kDa to 250kDa. The proteins are prestained with several fluorescent dyes and resolve into sharp, clearly identifiable bands for easy visualization and orientation. The green 10kDa, orange 70kDa and pink 140kDa bands serve as useful reference bands. HyperPAGE II is suitable to monitor migration efficiency in SDS-PAGE and protein transfer from gel to membrane in Western blotting, without the addition of staining dye. Prestained protein markers are suitable for determining the approximate molecular weights of proteins.
TRIsure provides a simple, efficient method for the sequential isolation of total RNA, genomic DNA and proteins from a wide variety of starting materials. By combining thiocyanate compounds with phenol, TRIsure facilitates disruption of cells during homogenization and effectively inhibits DNase and RNase activity. After homogenizing the sample with TRIsure, chloroform is added and the homogenate is allowed to separate into three phases: a clear aqueous phase (upper) containing the RNA, a green colored organic phase (lower) and an interphase, containing the DNA and proteins. The RNA is extracted from the aqueous phase by isopropyl alcohol precipitation, DNA is precipitated from the organic layer with ethanol and the proteins are precipitated from the phenol-ethanol supernatant by isopropyl alcohol precipitation. The precipitated RNA, DNA, or protein is washed to remove impurities and then resuspended for use in downstream applications. TRIsure allows for highly reliable integrative analysis of the transcriptome, genome and proteome, since the RNA, DNA and proteins are derived from the same sample and the same cell population. The simplicity of the TRIsure method allows simultaneous processing of a large number of samples and can be used in a wide variety of downstream applications including expression profiling, epigenetics, genotyping, transgenic analysis and cell line characterization.
Proteinase K is a highly active serine protease (MW 28,500 Da) isolated from the fungus Tritirachium album. The enzyme exhibits broad cleavage specificity on native and denatured proteins and is widely used in the purification of native RNA and DNA from tissues or cell lines. Because the solution is tested for the absence of RNases and DNases, it is ideal for isolating PCR and RT-PCR templates. The activity of Proteinase K is increased in the presence of denaturants such as SDS (1%) and elevated temperature (50-60°C). The recommended working concentration is 50-100 μg/mL for protein removal and enzyme inactivation and up to 2 mg/mL for tissue treatment. Proteinase K products are free of detectable DNase and RNase.
Product Highlights Excellent clarity - enhances visualization, even at high concentrations Strong gels at low concentration - better handling, less breakage DNase/RNase-free - for all nucleic acid separation Product Description Extremely pure, high molecular biology grade Agarose from Bioline has no detectable DNase or RNase activity and forms strong gels with low backgrounds. Due to its low EEO, DNA will have a high electrophoretic mobility.
Product Highlights Long oligonucleotide -more thermostable Reverse Transcriptases perform better with longer primers Binds only to mRNA -can be used for cDNA synthesis with total RNA Product Description Oligo (dT)18 Primer is suitable for use as a primer for first strand cDNA synthesis with a reverse transcriptase. The primer hybridizes to the poly-adenylated tail found on the 3´ end of most eukaryotic mRNAs. Oligo (dT)18 ensures that the 3´ end of mRNAs are represented. Primer sequence: 5´-d (TTT TTT TTT TTT TTT TTT)-3´ A mixture of random hexamer primers and oligo(dT) may improve the sensitivity of cDNA synthesis.
HyperLadder™ 50bp is a molecular weight marker, especially designed for easy size determination of linear double-stranded DNA fragments on 2% to 3% TAE or TBE agarose gels. The 10 regularly spaced bands, ranging from 50 bp to 2000 bp, with higher intensity reference bands at 300 bp, 1000 bp and 2000 bp allow for easy identification and orientation of your band at a glance. This ready-to-use format reduces handling steps and saves time; simply transfer HyperLadder 50bp from the vial to the gel. The concentration of DNA in each of the HyperLadder 50bp bands is provided for optionally determining the approximate mass of DNA in comparably intense sample bands of similar size. The HyperLadder 50bp is also supplied with a 5x loading dye for loading of sample DNA. HyperLadder 50bp is perfect for size determination in techniques such as confirmation of PCR products as well as other downstream techniques.
HyperLadder™ 25bp is a molecular weight marker, especially designed for easy size determination of linear double-stranded DNA fragments on 2% to 3% TAE or TBE agarose gels or polyacrylamide gels. The 10 regularly spaced bands, ranging from 25 bp to 500 bp, with higher intensity reference bands at 100 bp and 200 bp allow for easy identification and orientation of your band at a glance. This ready-to-use format reduces handling steps and saves time; simply transfer HyperLadder 25bp from the vial to the gel. The concentration of DNA in each of the HyperLadder 25bp bands is provided for optionally determining the approximate mass of DNA in comparably intense sample bands of similar size. The HyperLadder 25bp is also supplied with a 5x loading dye for loading of sample DNA. HyperLadder 25bp is perfect for size determination in techniques that produce very short fragments such as apoptotic DNA fragmentation as well as other downstream techniques.
MyTaq™ Red Mix is recommended for all standard PCR applications. MyTaq Red Mix is comprised of MyTaq DNA Polymerase and a novel buffer system that deliver very high yield PCR amplification over a wide range of PCR templates. MyTaq has an increased affinity for DNA, enabling reliable amplification from even very low amounts of template. MyTaq DNA Polymerase has been developed to give more robust amplification than other commonly-used polymerases allowing it to perform well with challenging templates and in the presence of PCR inhibitors. Furthermore, the highly efficient nature of MyTaq means it gives excellent results under fast PCR conditions. The product is supplied as a mastermix that requires the addition of only template, primers and water, thereby reducing the number of pipetting steps during PCR set-up, for improved speed, throughput and assay reproducibility. MyTaq Red Mix contains a red dye that increases the visual contrast between the reagent and the reaction vessel for improved convenience and to improve pipetting accuracy. The red dye also enables samples to be loaded directly on to a gel after the PCR without the need to add loading buffer. In addition, the MyTaq red reaction buffer contains dNTPs, MgCl2 and enhancers at optimal concentrations, which helps eliminate the need for optimization, thereby saving time, effort and the cost of performing unnecessary assay repeats.
MyTaq™ HS DNA Polymerase is recommended for PCR assays containing complex and low copy number targets as well as multiplex PCR. MyTaq HS DNA Polymerase is an antibody-mediated hot-start enzyme, specifically designed for highly-specific, efficient amplification from even the most challenging templates. MyTaq HS does not possess polymerase activity during the reaction set-up, thereby reducing the non-specific amplification that can hinder PCR assays from the start. The MyTaq HS DNA Polymerase and MyTaq Reaction Buffer in this product are a unique combination of next-generation hot-start polymerase and novel buffer system, that deliver very high yield PCR amplification over a wide range of PCR templates. MyTaq HS has an increased affinity for DNA, enabling reliable amplification from even very low amounts of template. MyTaq& HS DNA Polymerase has been developed to give more robust amplification than other commonly-used polymerases, meaning it performs reliably even in the presence of PCR inhibitors. Furthermore, the advanced formulation of MyTaq HS and buffer system allows fast cycling conditions, considerably reducing the reaction time without compromising PCR specificity or yield. MyTaq Reaction Buffer contains dNTPs, MgCl2 and enhancers at optimal concentrations, which helps eliminate the need for optimization, thereby saving time, effort and the cost of performing unnecessary assay repeats.
Product Highlights Accurate – clear discrimination of even the most challenging sequence differences Reproducible – unparalleled consistency between replicate difference plots for increased confidence in sample characterization Sensitive – efficient detection from even very limiting amounts of sample Flexible – enables reliable characterization of a broad range of sequence differences Fast – rapid PCR amplification prior to high resolution melting, enabling higher throughput "We used the Bioline SensiFAST HRM mix to discriminate changes at a locus with substitutions on both strands, yielding AT heteroduplexes as well as either A or T monoduplexes. We easily achieved calls on the mono duplexes over heteroduplexes with more than 99% confidence to provide extremely sensitive genotyping of an AT class 4 SNP." Dr Simon Baker, Senior Director of R&D, Bioline, London
Agarose gels are often run to check and see if a band is present or not, for example in a PCR reaction. With 5 regularly spaced bands, ranging from 100 bp to 2000 bp, EasyLadder™ I is a molecular weight marker especially designed for fast size determination of linear double-stranded DNA fragments on 0.5% to 3% TAE or TBE agarose gels. This ready-to-use format, with a red loading dye already added to the ladder, reduces handling steps and saves time; simply transfer EasyLadder I from the vial to the gel, along with the PCR product and run at high voltage for just a few minutes (1-3 cm of a gel lane). The concentration of DNA in each of the EasyLadder I bands is provided for optionally determining the approximate mass of DNA in comparably intense sample bands of similar size. The EasyLadder I is also supplied with a 5x loading dye for the loading of sample DNA.
RiboSafe RNase Inhibitor is a recombinant protein which completely inhibits a broad spectrum of eukaryotic RNases, including RNases A, B and C. Product Highlights Efficient – effectively inhibits a broad spectrum of eukaryotic RNases, including RNases A, B and C High-quality - certified free of contaminating DNases, RNases, Phosphatases and Nickases for improved sample security Robust – active up to 55 oC across a wide pH range and DTT concentrations, ideal for use most downstream applications Flexible – compatible with all common reverse transcriptases, bacterial polymerases and thermostable polymerases Product Description RiboSafe RNase Inhibitor is a recombinant protein which completely inhibits a broad spectrum of eukaryotic RNases, including RNases A, B and C. Furthermore, RiboSafe shows no inhibition of polymerase or reverse transcriptase activity and so can be used for cDNA synthesis and in one-step RT-PCR reactions. RiboSafe RNase Inhibitor is tested for activity, SDS-PAGE purity and the absence of endonucleases, nickases and exonucleases and functions robustly at a variety of temperatures and pH and in the presence or absence of DTT, allowing flexibility in designing experiments in highly-sensitive techniques such as single-cell RT-PCR, in vitro RNA synthesis and in vitro translation.
Product Highlights Mixture of oligonucleotides -for RNA missing a polyA tail such as bacterial RNA Binds randomly -for RNA with strong secondary structure Product Description Random Hexamer Primers consist of a mixture of oligonucleotides representing all possible hexamer sequences. Random Hexamer Primers are commonly used for priming single-stranded DNA or RNA for extension by DNA polymerases or reverse transcriptases. During cDNA generation, random priming gives random coverage to all regions of the RNA to generate a cDNA pool containing various lengths of cDNA. Random priming is incapable of distinguishing between mRNA and other RNA species present in the reaction. Primer Sequence: 5´ – d (NNNNNN) –3´ N = G, A, T or C A mixture of random hexamer primers and oligo(dT) may improve the sensitivity of cDNA synthesis.
MyTaq™ Red DNA Polymerase is recommended for all standard PCR applications. The MyTaq DNA Polymerase and MyTaq Red Reaction Buffer in this product, are a unique combination of next-generation polymerase and novel buffer system that deliver very high yield PCR amplification over a wide range of PCR templates. MyTaq has an increased affinity for DNA, enabling reliable amplification from even very low amounts of template. MyTaq DNA Polymerase has been developed to give more robust amplification than other commonly-used polymerases allowing it to perform well with challenging templates and in the presence of PCR inhibitors. Furthermore, the highly efficient nature of MyTaq means it gives excellent results under fast PCR conditions. The enzyme is supplied with a 5x MyTaq Red Reaction Buffer that increases the visual contrast between the reagent and the reaction vessel for improved convenience and to improve pipetting accuracy. The red dye also enables samples to be loaded directly on to a gel after the PCR without the need to add loading buffer. In addition, the MyTaq Red Reaction Buffer contains dNTPs, MgCl2 and enhancers at optimal concentrations, which helps eliminate the need for optimization, thereby saving time, effort and the cost of performing unnecessary assay repeats.
Product Highlights Ready-to-use - no need to add dye Dye visible in gel - allow users to monitor DNA migration Guaranteed batch-to-batch reproducibility - provides consistent results Product Description DNA Loading Buffer Blue is one of a range of Bioline Colored DNA Loading Buffers (fig. 1). The ready-to-use solution is premixed with bromophenol blue that migrate at different rates depending on the dye (fig.1 Lane 3) and the concentration of the agarose gel (see Dye Migration Table). This allow you to monitor DNA migration and therefore increase the versatility of your DNA analysis.
Highly quality dNTPs are a vital requirement for successful PCR, as the presence of contaminating impurities will result in a decrease in amplification sensitivity and product yield. Bioline ultra-pure dNTPs undergo highly stringent purification steps to give a greater than 99% purity and are tested or the absence of DNase, RNase, Protease, Nickase activity. They are then quality controlled& in a variety of applications, ensuring that they are highly suitable for the most sensitive of PCR techniques. All Bioline dNTP Mixes are supplied as an equimolar solution of ultra-pure dATP, dCTP, dGTP and dTTP as lithium salts. Lithium salts have greater resistance to repeated freezing and thawing cycles than sodium salts and remain sterile due to the bacteriostatic activity of lithium towards various microorganisms, giving greater reliability and an extended shelf life. As an ISO 13485 certified primary manufacturer of nucleotides, if your requirements for nucleotides are beyond the scope of our standard product range, Bioline can also offer custom, bulk and OEM services (firstname.lastname@example.org).