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.

highQu ALLin™ Taq DNA Polymerase is the versatile engineered enzyme which in combination with the optimized ALLin™ buffer provides higher success rates in demanding PCR applications like amplification of complex templates, crude sample PCR and fast cycling. ALLin™ Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 10^4 (a number of correct nucleotides incorporated before the error occurs) and produces A-tailed products suitable for ligating into TA cloning vectors. The convenience of ALLin™ Taq DNA Polymerase is maximized by the use of 2X Red Mastermix providing the additional advantage of reduced pipetting and minimized errors. ALLin™ Red Taq Mastermix, 2X is premixed with red dye and density reagents for direct loading on the gels after the PCR. In a 2% agarose TAE gel the dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments. The mastermix is even supplied with the PCR Water, and the only thing to add is the template with primers.

highQu ALLin™ HiFi DNA Polymerase is the outperforming high-fidelity enzyme derived from Pfu polymerase by introducing several point mutations. The robust engineered enzyme in combination with the optimized ALLin™ buffer provides higher fidelity (50X higher than Taq DNA Polymerase), better performance, increased success in demanding applications like amplification of complex or longer templates, crude samples and fast cycling. ALLin™ HiFi DNA Polymerase produces blunt-ended products suitable for ligating into blunt vectors.

highQu ready to use DNA ladders are mixtures of highly purified plasmid digests and PCR products. The high ladder purity allows for exceptional stability and room temperature storage. Ladders are ready to be directly loaded on agarose gels and are supplied with the loading dye solution for probe DNA. They provide sharp bands for DNA sizing and allow for approximate DNA quantification by comparing the amount of DNA of the band similar in size to your sample band. The most important factors influencing agarose gel electrophoresis results:  Use the right percentage of the agarose gel for the given size of the DNA to be analysed. Prepare the gel according to the instructions of the agarose manufacturer. Use the most sensitive nucleic acid stain (StainIN™ Green) in both agarose gel and in the buffer. Use the same loading dye both for the samples and for the DNA ladder. Select DNA ladder which is supplied with the loading dye solution for the DNA samples (Take5™ 1 kb DNA Ladder). For more precise sizing of the DNA, load the DNA ladder into the two slots of the agarose gel Use enough of the electrophoresis buffer to keep the agarose gel well-covered during the electrophoresis run.  Visualize the agarose gel to compare your sample bands with the ones of the DNA ladder using an optimal gel documentation system.