EasySep™ Total Nucleic Acid Extraction Kit

总览

从全血、细胞悬液（如PBMCs、hPSCs、培养过的细胞）和EasySep™分选出的细胞中高效提取总核酸（DNA和RNA）。EasySep™总核酸提取试剂盒使用简单，可用于大通量提取，采用磁珠技术，可以避免使用离心柱或有毒试剂。该试剂盒提取的核酸纯度高，可直接用于下游应用，如qPCR。

使用本试剂盒用EasySep™total nucleic Acid RapidSpheres™对样品中的总核酸进行磁性标记。然后，用磁铁（单独出售）处理样品——无需使用离心柱。使用移液管弃去未标记的非目标成分后，再从磁铁中取出样本，得到标记的核酸。可使用1.7mL微量离心管搭配ErythroClear™磁铁（产品号#01737）；或为了扩大提取通量，使用96孔PCR微孔板搭配96孔PCR微孔板磁铁（产品号#100-1304）在96孔PCR板中处理样品。

磁体兼容性
ErythroClear™ Magnet (Catalog #01737) 96-Well PCR Microplate Magnet (Catalog #100-1304)

细胞类型
淋巴细胞，单个核细胞

应用
基因组编辑，核酸纯化

研究领域
癌症，免疫

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实验数据

Diagram of the EasySep™ Total Nucleic Acid Extraction Kit workflow.

Figure 1. EasySep™ Total Nucleic Acid Extraction Kit Workflow

Diagram of the standard extraction workflow. Time points at which samples must be placed in the magnet are indicated with gray boxes. EasySep™ Lysis Buffer and Proteinase K are added to the sample and incubated at 56°C for 10 minutes. Diluted EasySep™ Nucleic Acid RapidSpheres™ are added to the sample and incubated at room temperature (RT) for 5 minutes, then placed in the magnet for 2 minutes. While in the magnet, the sample is washed three times with a 70% ethanol wash solution, and the supernatant is removed. The pellet is resuspended with the elution buffer while removed from the magnet and incubated at room temperature for 5 minutes. The sample is then placed in the magnet for 2 minutes before the supernatant is aspirated to a new tube to obtain the extracted nucleic acids.

Figure 2. EasySep™ Total Nucleic Acid Extraction Kit Shows Improved DNA and RNA Recovery Relative to Other Commonly Used Methods

Normalized recovery (μg per 1 x 10^6 cells) of (A) DNA and (B) RNA across EasySep™ Total Nucleic Acid Extraction Kit and other commonly used extraction methods. DNA and RNA concentrations were measured separately using the Qubit™ Fluorometer. Sample source: Leukopak, n = 3. Error bars represent ± 1 standard deviation.

EasySep™ Total Nucleic Acid Extraction Kit recovers nucleic acids of optimal purity when benchmarked against other commonly used extraction methods.

Figure 3. EasySep™ Total Nucleic Acid Extraction Kit Recovers Nucleic Acid of Optimal Purity

Purity ratios obtained using spectrophotometric absorbance measurements show that the nucleic acid recovered by the EasySep™ Total Nucleic Acid Extraction Kit is of optimal purity, benchmarked against other commonly used extraction methods. The 260/280 ratio, indicative of protein, phenol, and other contaminants within the extract that absorb at or near 280 nm, has an optimum purity ratio of ~ 1.8 for DNA and ~ 2.0 for RNA. The 260/230 ratio, a secondary measure of purity indicative of salt, phenol, and other organic compound contamination, has an optimal range of 1.8 - 2.2. Ratios that fall outside of these ranges are generally considered contaminated. Purity ratios were obtained using the Nanodrop™ Spectrophotometer. Sample source: Leukopak, n = 3. Error bars represent ± 1 standard deviation.

Results from a DNA-based qPCR assay with primers targeted to a non-genic region of chromosome 4 demonstrate that, DNA is detectable in extractions with a starting cell input of 10 cells and up to 1 x 10^6 cells, using the EasySep™ Total Nucleic Acid Extraction Kit.

Figure 4. EasySep™ Total Nucleic Acid Extraction Kit Is Capable of Extracting from As Few As 10 Cells

Results from a DNA-based qPCR assay using primers targeted to a non-genic region of chromosome 4 demonstrate that DNA is detectable in extractions with a starting cell input of 10 - 1 x 10^6 cells. (A) qPCR curves coloured by cell input. Dotted red line represents the cycle threshold (Ct; 0.139). Each individual curve represents 1 technical replicate, n = 3. (B) Inverse linear relationship between log(cell number input) and Ct value. Each data point represents 1 technical replicate, n = 3. Sample source: Leukopak.