highQu PCRbeam™ Fast PCR Detection Kit is a convenient tool for fast detection of gene-specific amplification products obtained by PCR, LAMP or RPA. The detection is based on immunological reaction driven by Biotin and FITC (fluorescein isothiocyanate), thus the amplified DNA shall include Biotin and FITC labels. The PCR amplification has to be performed with one primer labeled with FITC at 5’-end and one primer labeled with Biotin at 5’-end. Alternatively the use of one of the labeled primers can be replaced by gene-specific FITC or Biotin labeled probe. Kit includes PCRbeam™ Membrane Strips that are coated with biotin-ligand on the test band and an anti-rabbit antibody on the control band. The bottom part of the strip which is used for sample application contains an anti-FITC antibody attached to gold particles. PCRbeam™ Detection Buffer is Tris-buffered saline enabling the detection. The PCRbeam™ Fast PCR Detection Kit can be applied for established tests or home-brew assays as a fast and sensitive yes/no detection method. The detection sensitivity is up to 100 fold higher than the one achievable with ethidium bromide stained gels what provides an environment friendly save and economical alternative to the use of mutagen stains. For establishing sensitive PCR-based tests before PCRbeam™ detection we recommend the use of hot-start PCR enzymes or master mixes, like highQu ALLin™ Hot Start Taq Mastermix or ALLin™ Hot Start Taq DNA Polymerase. Principle: The membrane strip is soaked for 10 minutes into the vial with the detection buffer mixed with PCR product. The lateral sample flow driven by gold particles moves the solution up the strip. FITC labeled DNA strand binds with the anti-FITC antibody on the gold particle and Biotin labeled DNA strand is caught by Biotin ligand attached to the test band. As both DNA strands remain hybridized at room temperature, the test band builds an aggregate that develops red-blue color. Excess gold particles that were not caught by FITC move up the strip and the anti-FITC antibody binds to the anti-rabbit antibody to develop the red-blue colored control band. If there is no PCR product in the reaction, then only the control band will be visible. If there is a specific product, the test band will be colored as well.

Derived from our HiFi polymerase, the highQu ALLin™ Mega HiFi DNA Polymerase provides much lower error rate PCR with a 100 higher fidelity compared to Taq. The ALLin™ Mega HiFi DNA Polymerase is engineered to be much faster and to generate a higher yield of long PCR products up to 20 kb from complex GC-rich templates. Therefore the ALLin™ Mega HiFi DNA Polymerase is an excellent choice for longer and very complex PCR applications where the highest fidelity is demanded. It is an enzyme of choice for cloning and all kind of sequencing applications including NGS. Generated blunt-ended PCR products are suitable for ligation into blunt vectors. For maximum convenience, the ALLin™ Mega HiFi Mastermix, 2X (HLM0201) and ALLin™ Mega HiFi Red Mastermix, 2X (HLM0301) are available. Pfu DNA Polymerase was originally found in thermophilic bacterium Pyrococcus furiosus. It is a thermostable DNA polymerase that was widely used in molecular biology applications for its high fidelity of DNA amplification due to a high proofreading activity. Pfu polymerase has 3' to 5' exonuclease proofreading activity, which enables it to correct errors that occur during DNA synthesis. The enzyme is commonly used for cloning and sequencing applications, however it is relatively slow, sensitive to contaminants and inhibitors and has low processivity resulting in usually low PCR yields. Alternative, mostly better DNA polymerases have been developed by different companies to address the limitations of Pfu polymerase. These include such enzymes as Phusion® DNA polymerase, Q5® DNA polymerase from NEB, and many others. These enzymes offer improved elongation rates, reduced error rates, and increased efficiency for NGS applications. The ALLin™ MEGA HiFi Polymerase offered by highQu provides 100X higher fidelity compared to Taq Polymerase, which is much higher than Pfu. In addition, the hot start enzyme version allows to reduce the background to the minimum. Such robust enzyme offers not only precise amplification, but also enables PCR from very complex templates and generation of very long PCR products up to 30 kb and above. Availability of faster, more robust and more precise polymerases influenced successful development of next generation sequencing techniques (NGS) where PCR precision and yield is crucial for reliable sequencing results.

highQu ALLin™ RPH Polymerase (Robust, Proofreading, Hot-start Polymerase) is the versatile engineered enzyme combining best polymerase properties for excellence in most demanding PCR applications, like low copy detection, long or high fidelity PCR, amplification of complex templates, crude sample PCR and multiplexing. ALLin™ RPH Polymerase has 5 times higher fidelity than Taq DNA Polymerase and produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the ALLin™ RPH Mastermix, 2X is available.

highQu Synthetic Carrier RNA is designed to be used in all kind nucleic acid purification and precipitation procedures as a carrier and co-precipitant of nucleic acids. It is especially useful to increase the amount of RNA or DNA pellet in low concentrated solutions, in such procedures, as viral RNA extraction from human specimen samples. In contrast to commonly used carrier RNAs such as tRNA, yeast RNA, or sonicated salmon sperm DNA, the Synthetic Carrier RNA is free from animal or yeast RNA contamination. Coprecipitated RNA and DNA can be directly used for all common downstream applications, such as PCR or RT-PCR, as well as highly sensitive qPCR. The use of carrier RNAs for coprecipitation of nucleic acids may interfere with spectrophotometrical concentration measurements. The presence of carrier RNAs in the RNA or DNA solution may have some influence on certain enzymatic reactions performed by such enzymes that act on all nucleic acid molecules, for example T4 Polynucleotide Kinase or Terminal DNA Transferase.

highQu PCR Water is a supplementary high quality reagent for all demanding PCR and qPCR and other molecular biology applications. It saves time being on your bench and guaranties the purity of reactions and inhibition-free performance of PCR reagents. highQu PCR Water is a deionized, membrane filtered water continuously tested in ultrasensitive qPCR and PCR applications, in amplification of long targets and highly specific detection of few copies of templates.

Proteinase K (Molecular Biology Grade Solution) is a serine peptidase with a very high specific activity and a broad spectrum of protein digestion possibilities. The solution is designed to be used for protein degradation (up to tetrapeptides) during the cell lysis and RNA/DNA extraction procedures under hush reaction conditions such as the higher temperatures and the presence of detergents. The enzyme efficiently degrades DNases and RNases during nucleic acid isolation process. The high purity of the Proteinase K and controlled absence of both DNAse and RNase contamination ensures the integrity of nucleic acids. The 100% enzyme activity (when stored at -20°C) is guaranteed for at least two years after production. However, the experiments proved that the proteinase remains close to 90% active even when stored at +37°C for 18 months. The enzyme is supplied as a 20 mg/ml concentrated solution with an average specific activity of more than 800 u/ml. Active in all common buffers used for cell lysis and RNA/DNA extraction, in a presence of urea, SDS and guanidinium salts Stable at high temperature of up to 56°C Can be inactivated by heating at 65oC for 20 minutes or at 75oC for 10 minutes Active in a pH range of 4–12 with an optimum pH 7.5–8.0 The Proteinase K gene from album expressed in yeast host. The quality limit of allowed host DNA presence is ≤ 0.25 pg/U measured by qPCR what is about a half when compared to other suppliers. Unit Definition Folin & Ciocalteu’s method - One unit is required to hydrolyze urea-denaturated hemoglobin producing color equivalent of 1 μmol tyrosine in 1 minute at 37°C and pH 7.5, 1 U = 1 m Anson U.