Multi PCR Enhancer

Item Number

M748274

• Specifications & Purity: 2X
• Stability And Storage: Store at -20℃.
• Storage Temp: Store at -20°C
• Shipped In: Ice chest + Ice pads

Product Description

Aladdin's Mult PCR Enhancer (2X) is a PCR enhancer designed to overcome the barriers to PCR amplification caused by the high GC content of templates when using Mult Multiplex PCR Kit . The amplification of DNA fragments with high GC content has been one of the difficulties in molecular biology, and this product can be used in direct PCR reactions containing anticoagulants such as heparin, sodium citrate, and EDTA, to improve the amplification efficiency and specificity of DNA templates with high GC content. It can also be used in routine PCR to amplify target fragments with high GC content.The Mult PCR Enhancer (2X) is compatible with a variety of commercial blood-resistant DNA polymerases, such as Aladdin's EnzymoPure™ DNA Polymerase (Blood-resistant) , EnzymoPure™ HF DNA Polymerase (Blood-resistant) , and NEB's Hemo KlenTaq, etc.This product is significantly better than betaine and DMSO in improving the PCR amplification of GC-rich genes from EDTA anticoagulated blood.This product is sufficient for 80 PCR reactions of 50μl or 200 PCR reactions of 20μl .

Precautions：

The Multi™ PCR Enhancer (2X) must be used in conjunction with the 1X PCR Buffer included in Multi™ Multiplex PCR Kit.If crystalline precipitates are present in this product after thawing, incubate it in a 50℃ water bath for a few moments to dissolve the precipitates completely before use.We recommend using the freshly collected anticoagulated blood in PCR reactions. Repeated freeze-thaw of frozen samples that may cause fragmentation and degradation of target genes should be avoided.The anticoagulated blood can be added directly to the PCR reaction without any additional processing, and the recommended amount is about 5-10% of the total volume of the PCR reaction.After amplification, centrifuge the PCR reaction mix at 3000-5000×g for 3-5min to precipitate the blood cell fractions and then take the supernatant for electrophoresis. It is normal to have transparent gelatinous precipitations at the bottom of the PCR reactions because of the presence of anticoagulants.Because the PCR reaction is extremely sensitive, contamination must be avoided during preparation of the PCR reactions. Negative control without templates is recommended for all PCR assays to control contamination.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Please follow the instructions of Multiplex PCR Kit to set up PCR running conditions. Usually, for amplification of GC-rich DNA templates, the extension time should be longer than the conventional extension time.2. Set up the PCR reaction as follows:a. Thaw the reagents required for PCR reaction setup. Mix well and keep the DNA polymerase on ice until use.b. Set up the PCR reaction on ice. When multiple PCR reactions are required, a master mix can be prepared and dispensed into each PCR reaction tube.ComponentVolume (μl)Volume (μl)Final Nuclease-free Water(5-x-0.4n)μl(12.5-x-n)μl-10X PCR Buffer2μl5μl1X PCR Enhancer (2X)10μl25μl-dNTP (2.5mM each)2μl5μl0.25mM eachPrimer Mix (10μM each)0.4μl×n1μl×n0.2μM eachTemplatexμlxμl-Control Template and Primer Mix---Multiplex™ DNA Polymerase1μl2.5μl-Total Volume20μl50μl-Note 1: n represents the number of primer pairs used in multiplex PCR reaction (i.e., the number of target DNA).Note 2: The amount of DNA template has significant influence on PCR amplification. For high complexity DNA samples, such as mammalian genomic DNA, it is recommended to use 5ng to 0.5μg in a 20μl PCR reaction. For low complexity DNA, such as lambda DNA or plasmid DNA, it is recommended to use 5pg to 5ng in a 20μl reaction.Note 3: For blood samples: The amount of anticoagulated blood used in PCR reactions is generally 1-20% of the total volume, and the recommended starting amount is 5%. For detecting genomic DNA fragment in blood, lower blood concentration can be used. To detect the viral, bacterial or other microbial DNA in blood samples, it is recommended to use a 50μl PCR reaction and use a larger amount of blood. For amplification of GC-rich DNA fragments, either add the PCR Enhancer (2X) or supplement 1-10% DMSO in PCR reactions. Note 4: For plant samples: The recommended plant sample for 20μl and 50μl PCR reactions is 0.1-1mm and 0.3-3mm diameter leaves or other tender plant tissues of similar size, respectively. Fresh and tender seeds can be added directly in PCR reactions, but not dry and hards seeds. Use a clean scalpel to remove the seed shell, cut out 0.5-2mm diameter tissue and put it directly into the PCR reacion. One to two small seeds such as tomato seeds, can be added directly into PCR reactions for DNA amplification. In 50μl of PCR reactions, the diameter of plant leaf or seed should not exceed 3mm, as too many PCR inhibitors present in plant samples will inhibit PCR amplification. It is recommended to perform two parallel PCR reactions for the same sample to reduce variations of sampling. To ensure the uniformity of sampling, it is recommended to use a specialized punch or scalpel for sampling, and to prevent cross-contamination by cleaning the punch or scalpel with 2% sodium hypochlorite solution between each sampling. To amplify GC-rich DNA fragments, supplement 1-10% DMSO in PCR reaction can be attempted.Note 5: The final concentration of each primer in PCR reactions is recommended to be 0.2μM, but can be adjusted in the range of 0.05-0.4μM. Increase the primer concentration when amplification efficiency is low and decrease the primer concentration when non-specific PCR products are amplified.c. Mix well gently by vortex or pipetting. Centrifuge briefly at room temperature to collect the liquid at the bottom of the tube.d. (Optional) When using a thermocycler without a hot lid, add a drop of mineral oil to the reaction to avoid evaporation. e. Transfer the PCR reaction to a thermal cycler and run thermocycling conditions as recommended for the Multiplex PCR Kit. The extension time can be prolonged.Related Products:产品编号产品名称包装D7241MHemoTaq™ DNA Polymerase (Blood-resistant)1000TD7241SHemoTaq™ DNA Polymerase (Blood-resistant)200TD7243MHemoTaq™ HF DNA Polymerase (Blood-resistant)1000TD7243SHemoTaq™ HF DNA Polymerase (Blood-resistant)200TD7245Blood PCR Enhancer (2X)2mlD7248MPlantTaq™ DNA Polymerase (Chlorophyll-resistant)1000TD7248SPlantTaq™ DNA Polymerase (Chlorophyll-resistant)200TD7285MEasy-Load™ Blood Direct PCR Master Mix (2X)500TD7285SEasy-Load™ Blood Direct PCR Master Mix (2X)100TD7287MEasy-Load™ Blood Direct PCR Master Mix (HF, 2X)500TD7287SEasy-Load™ Blood Direct PCR Master Mix (HF, 2X)100TD7289MEasy-Load™ Plant Direct PCR Master Mix (2X)500TD7289SEasy-Load™ Plant Direct PCR Master Mix (2X)100TD7301M Multiplex PCR Kit500D7301S Multiplex PCR Kit100D7303S PCR Enhancer (2X)2mlD7305MEasy-Load™ Multiplex PCR Master Mix (2X)500D7305SEasy-Load™ Multiplex PCR Master Mix (2X)100