The qScriber™ cDNA Synthesis Kit is a highly efficient and simple-to-use system for cDNA synthesis eliminating the need for tedious reaction optimization. The qScriber™ Enzyme Blend ensures high sensitivity detection from low copy number targets. The highly active and thermostable HighScriber™ Reverse Transcriptase blended with RNase Inhibitor allows for an efficient cDNA synthesis and reaction safety. The wide reaction temperature range (38°C - 55°C) ensures efficient transcription from GC rich templates. The 5X qScriber™ Reaction Mix includes optimal concentrations of magnesium and dNTPs and a combination of anchored oligo (dT) and random hexamers for unbiased representation of mRNA ends. The kit is an optimal choice for generating high quality cDNA from viral RNA, miRNA or other targets for qPCR or for PCR. For the maximum convenience the Kit includes even the PCR Water to set up the reaction, so the only thing you need to take care of is the high quality RNA template.   cDNA (complementary or copy-DNA) synthesis is a fundamental technique used in both molecular biology research and in diagnostic applications. Reverse transcription process employs reverse transcriptase enzymes (RT), RNase inhibitors, template, random hexamer, oligo dT or specific primers and dNTPs. cDNA is produced aiming to perform different downstream applications. For example, for cloning purposes one takes specific primers, and performs the PCR after the cDNA synthesis to generate long DNA fragments for cloning into the plasmid vectors and for sequencing. For gene expression analysis purposes, short random primers are used, and generated short cDNA fragments unbiased represent the whole genome for qPCR analysis. A good cDNA synthesis kit provides high yields of high-quality cDNA with minimal bias or variability. For complex targets containing high-GC regions thermostable reverse transcriptases are preferred, as they allow performing the reaction at higher temperature what prevents formation of secondary structures and increases the cDNA yield. Typically, the optimal cDNA synthesis temperature ranges between 37-42°C, depending on the reverse transcriptase enzyme used. Advanced RT enzymes go up to 55°C and above. Optimizing cDNA synthesis protocol is crucial for generating accurate cDNA libraries. Several factors can influence the quality and yield of cDNA, including the amount and quality of RNA template, the concentration and quality of reverse transcriptase enzyme, quality of the primers, and the cDNA synthesis temperature. At highQu, we offer several kits and enzymes for cDNA synthesis applications, these include: 1Step RT PCR Kit for fast PCR, HighScriber™ Reverse Transcriptase Mix for cloning purposes, and qScriber™ cDNA Synthesis Kit for a real-time qPCR-based gene expression analysis.

highQu 1Step RT PCR Kit combines the 20X RT and RNase Inhibitor mix for efficient reverse transcription and the 2X PCR Mastermix for subsequent amplification of cDNA in the same tube. RT2 Mix, 20X is a blend of the engineered MMuLV (stable at 40-55°C allowing for high yields of long transcripts) with an efficient Ribonuclease Inhibitor protecting the template RNA from RNases. The resulting cDNA is then amplified by the 1Step RT PCR Mastermix, 2X. The PCR mastermix contains our proprietary Hot Start Taq DNA Polymerase. The enzyme activity at room temperature is blocked by small molecular inhibitor. Enzyme becomes active only after heating what allows for highly specific and extremely sensitive amplification, no primer dimer formation and no background. In combination with the optimized buffer the enzyme provides higher success rates in demanding PCR applications like amplification of complex or longer templates and fast cycling. Hot Start Taq DNA Polymerase has the same PCR accuracy like Taq DNA Polymerase, 4.5 x 104 (nucleotides incorporated before the error occurs), and produces A-tailed products suitable for ligating into TA cloning vectors. For the maximum convenience the Kit includes even the PCR Water to set up the reaction, so the only thing you need to take care of is the high quality RNA template.

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.

  StainIN™ eco-RED Nucleic Acid Stain, a 10000X concentrated aqueous solution is a significantly safer alternative to ethidium bromide. It is same easy to use, twice as sensitive and much more secure. At least twice as economical as competing products, this novel DMSO-free stain can be also used and disposed with less environmental and health concerns compared to ethidium bromide. StainIN™ eco-RED is a fluorescent dye that allows detection of >0,1 ng of DNA in both agarose and polyacrylamide gels. It binds to both ds DNA, ssDNA and RNA and emits red fluorescence detectable under the LED or UV light and documented with same filters as ethidium bromide. For cloning applications, LED light is recommended. The lower than ethidium bromide carcinogenicity of the dye has been proven by Ames-test.    The carcinogenicity and a high toxicity of a widely used gel stain Ethidium Bromide (EtBr) is one of the biggest safety and environmental problems in the labs where gel electrophoresis is a daily routine. Therefore, there is a need for a safer ethidium bromide alternatives to be used in laboratories. The novel less toxic nucleic acid dyes used for agarose gel electrophoresis address this ethidium bromide problem and minimize environmental concerns. highQu StainIN™ eco-RED Nucleic Acid Stain provides a significantly safer alternative to ethidium bromide while delivering up to several times higher detection sensitivity. Same like ethidium bromide, the stain is used during the process of electrophoresis in an agarose gel or PAAG and (optionally) in the electrophoresis running buffer, or alternatively, for a post-run staining of the gels in a tray with the buffer. Despite having a proven lower mutagenicity, all nucleic acid stains shall be handled with care, as they interact with DNA and RNA.  Used electrophoresis buffer with a stain as well as melted gels shall be run through approved (carbon) filters. If the absence of residual fluorescence is confirmed, the liquids can be disposed with plenty of water down the drain. Alternatively, all used liquids can be autoclaved or heated as the StainIN™ eco-RED Nucleic Acid Stain degrades during the long exposure to above 100ºC temperature. The safety office shall be consulted periodically to match local regulations, as these regulations vary and change. Despite of tests described in this report, the seller of the listed stains and other reagents does not take any responsibility for the possible damage resulting from handling any sold chemical or reagent.   Frequently asked questions about the new Nucleic Acids Stain 1.      What are the advantages of the new StainIN™ eco-RED NA Stain over the old version StainIN™ RED? The new stain is purely aqueous solution. The old StainIN™ RED had DMSO as solvent.  The new stain is room temperature stable, shipped at ambient temperature. Store it at your electrophoresis bench. The old StainIN™ RED was stored at +4°C.  StainIN™ eco-RED NA Stain is more flexible – detectable under both UV and LED light. More convenient protocol – the stain can be used both during the gel run and as a post-run staining. Same or better sensitivity compared to the old stain. 2.      Is the color of the solution the same as before?  The new stain is a darker shade of red. 3.      What is the concentration of the StainIN™ eco-RED Nucleic Acids Stain?  It is 10,000X concentrated, however the number of applications for post run staining is difficult to calculate because multiple gels can be stained in the same staining solution.  4.      What is the solvent used in the StainIN™ eco-RED Nucleic Acids Stain? The solvent is 100% water. 5.      Which stains are similar to StainIN™ eco-RED Nucleic Acids Stain?  The closest similar stains are Ethidium Bromide and GelRed (Biotium). 6.      What is the sensitivity of the StainIN™ eco-RED Nucleic Acids Stain in ng/band?  The sensitivity ranges from 0.1 to 0.3 ng DNA/band or even higher, depending on factors such as the staining method used, imaging capability, detection, and gel thickness. 7.      What is the emission wavelength of the StainIN™ eco-RED Nucleic Acids Stain?  The emission wavelength is 600 nm. 8.      What is the excitation wavelength of the StainIN™ eco-RED Nucleic Acids Stain?  The excitation wavelength is 300 nm. 9.      Can StainIN™ eco-RED Nucleic Acids Stain be used for in-gel and in-buffer staining?  Yes, it can be used for both in-gel and in-buffer staining post electrophoresis run. 10.   How should the stain be used during a gel run in agarose gels?  Use 8-10 µl of the stain in 100 ml agarose gel solution cooled down for pouring, nothing to be added in to the electrophoresis buffer. 11.   How to use StainIN™ eco-RED Nucleic Acids Stain with PAAGE gels? Same like with agarose gel, or using the post-run staining protocol. 12.   Can the StainIN™ eco-RED Nucleic Acids Stain be detected under UV light?  Yes, it can be detected under UV light, with ethidium bromide or GeRed filters. 13.   What color does the stained DNA appear as under UV light?  DNA appears as red-orange under the UV light. 14.   What color does RNA appear as under UV light?  RNA appears as red-orange under UV light. 15.   Does StainIN™ eco-RED Nucleic Acids Stain color both single-stranded and double-stranded DNA?  Yes, it can stain both single-stranded and double-stranded DNA, and RNA. 16.   Can the stained DNA be detected with a blue LED?  Yes, the StainIN™ eco-RED Nucleic Acids Stain -stained DNA can be detected with a blue LED. 17.   Is StainIN™ eco-RED Nucleic Acids Stain compatible with cloning?  Yes, as it can be used with LED without damaging DNA. 18.   Has StainIN™ eco-RED Nucleic Acids Stain undergone toxicity testing? Yes, it is less toxic than Ethidium Bromide See the toxicity testing report. 19.   Can StainIN™ eco-RED Nucleic Acids Stain be used for post-run staining?  Yes, it can be used for post-run staining. 20.   Can the staining buffer (electrophoresis buffer with the NA stain) be reused?  Yes, the staining buffer can be reused several times, depending on gel thickness and the amount of DNA stained. It should be stored in the dark in a glass or plastic container. To avoid StainIN™ eco-RED Nucleic Acids Stain absorbance on plastic or glass over the prolonged storage of more than 24 hours, make a fresh staining solution every day. 21.   Is the StainIN™ eco-RED Nucleic Acids Stain sensitive to light?  Yes, same as all other fluorescent nucleic acid’s stains, the StainIN™ eco-RED Nucleic Acids Stain is light-sensitive and should be protected from light. 22.   Can StainIN™ eco-RED Nucleic Acids Stain be stored at +4°C?  The recommended storage conditions for this stain is ambient temperature at about +20°C.  However, due to common cooled shipments, it can be exposed for a short time to +4°C. If accidentally shipped with cooling packs at +4°C, simply mix the product to ensure the stain is completely dissolved. And keep it further only at ambient temperature. 23.   What happens if the StainIN™ eco-RED Nucleic Acids Stain was put or exposed to -20°C and was frozen?  If frozen, the StainIN™ eco-RED Nucleic Acids Stain should be disposed and the new product shall be ordered. Do not use it if once frozen.