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.

highQu Taq DNA Polymerase is the classical enzyme for routine PCR applications providing high amplification yields of 3-5 kb targets under various conditions. Taq DNA Polymerase is purified from a recombinant E. coli strain carrying the Taq DNA polymerase gene. Taq DNA polymerase is thermostable 5´ → 3´ DNA polymerase. It lacks 3´ → 5´ exonuclease (proofreading) activity and has low 5´ → 3´ exonuclease activity. Polymerase exhibits deoxynucleotidyl transferase activity resulting in A-overhang at the 3'-ends of PCR products and allowing for TA cloning. The PCR accuracy of Taq DNA Polymerase is 4.5 x 104 (a number of incorporated nucleotides before the first error occurs). The supplied reaction buffer for most flexibility in optimization, contains no dNTPs and no magnesium. The 50 mM MgCl2 solution is provided separately what allows for magnesium optimization starting from 1 mM up to 4 mM final concentration upon the need. The dNTPs in sets or mixes can be purchased from highQu separately. The most widely used PCR enzyme is the DNA polymerase from Thermus aquaticus. Taq DNA polymerase amplifies template DNA by adding nucleotides to the 3' end of a DNA strand in a template-dependent manner where short single-stranded primers that anneal to the template provide a starting point for polymerization. The enzyme has a 5' - 3' polymerase activity and lacks 3' - 5' exonuclease or so-called proofreading activity. For optimal performance, Taq DNA polymerase requires not only template DNA and reverse and forward primers, but also such essential PCR reaction components as deoxyribonucleotide triphosphates (dNTPs), and magnesium chloride. Optimized buffers commonly supplied with commercial Taq Polymerase enzymes contain certain salt concentration and have balanced pH value. Taq DNA polymerase has several other advantageous properties over other enzymes used for the PCR. It is known for a high stability and resistance to PCR inhibitors. The inhibitor resistance allows performing direct PCR reactions from crude non-purified templates, such as colony PCR. At highQu, Taq DNA Polymerase is either available as a stand-alone enzyme supplied with a ALLin™ Buffer, or as a convenient 2X concentrated master mix formulation.

highQu ALLin™ Mega HS HiFi DNA Polymerase provides much lower error rate PCR with a 100 higher fidelity compared to Taq. Compared to Mega HiFi, this hot start enzyme version allows for even higher sensitivity and specificity of PCR as well as for a room temperature reaction setup, and is excellent choice for multiplex reactions. The ALLin™ Mega HS HiFi DNA Polymerase is engineered to be much faster and to generate higher yield of long PCR products up to 20 kb from complex GC-rich templates. Therefore the ALLin™ Mega HS 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 HS HiFi Mastermix, 2X (HLM0401) and ALLin™ Mega HS HiFi Red Mastermix, 2X (HLM0501) are available.

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.