Blood PCR Enhancer

Item Number

B744167

• 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 Blood PCR Enhancer (2X) is designed to improve the amplification of DNA with high GC content. It can also be used for direct PCR of anticoagulated blood containing heparin, sodium citrate, or EDTA, to significantly improve the amplification efficiency and specificity.This product 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.Please refer to Figure 1 for the effect of this product on the direct amplification of genes with high GC content in EDTA-anticoagulated blood.Figure 1. GC-rich DNA fragements amplified in PCR reactions containing Aladdin's Blood PCR Enhancer (2X) . A. HRES1 fragment (488bp, 81% GC content) and DIP2A fragment (1185bp, 71% GC content) can be amplified after the addition of this product, but cannot be amplified without this product; B. Compared to similar products from Company A and Company B, Aladdin's Blood PCR Enhancer (2X) has better performance in amplifying the 488bp fragment of HRES1 gene, and better than 1.3M Betaine as well.When a PCR reaction volume of 50μl is used, this product is sufficient for 80 reactions. When a reaction volume of 20µl is used, this product is sufficient for 200 reactions.

Precautions：

The Blood PCR Enhancer (2X) must be used in conjunction with the 1X PCR Buffer that is compatible with the blood-resistant DNA polymerase.If crystalline precipitates are present in this product after thawing, incubate in a 50℃ water bath for a few moments to dissolve the precipitates completely before use.We recommend using freshly collected anticoagulated blood for PCR. 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：

PCR reaction setup:a. Thaw PCR components at room temperature and mix well prior to use. Keep the HemoTaq™ DNA Polymerase on ice.b. Set up the reaction in a PCR tube on ice as follows:ComponentVolume (μl)Volume(μl)Final Nuclease-free Water(14-x) μl(35-y) μl-10X HemoTaq™ PCR Buffer2μl5μl1XdNTP (2.5mM each)2μl5μl0.25mM eachControl Primer Mix (10μM each)1μl2.5μl0.5μM eachHemoTaq™ DNA Polymerase1μl2.5μl-Anticoagulant Whole Bloodxμlyμl-Total Volume20μl50μl-Note 1: When multiple reactions are required, prepare a master mix including all reagents except template and primer, and then dispense to different PCR tubes. Sometimes, the master mix can also include template and primer.Note 2: The final concentration of each primer is recommended to be 0.2μM, but can be adjusted in the range of 0.1-1.0μM. It can be increased when the amplification efficiency is not high, and can be reduced when non-specific amplification occurs.Note 3: The amount of anticoagulated blood used as PCR template is generally 1-20% of the total volume of the PCR reaction, 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 PCR amplification of GC-rich DNA, either add the Blood PCR Enhancer (2X) ( 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)100T