PAXgene Blood DNA Kit for Genomic DNA Isolation

从全血样本中纯化至多500 μg基因组DNA

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PAXgene Blood DNA Kit (25)

目录编号 / ID. 761133

Processing tubes and buffers for 25 preparations

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仅作研究用途。不用于诊断程序。任何表述或表示都并非旨在提供诊断、预防或治疗疾病的信息。

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特点

同一体系中进行血液收集和纯化
易于操作
提高工作流程的效率

产品详情

PAXgene Blood DNA系统包括用于血液采集和稳定的PAXgene Blood DNA Tubes和用于单管纯化DNA的PAXgene Blood DNA Kit。

绩效

PAXgene Blood DNA System分离的DNA A₂₆₀/A₂₈₀比值在1.7–1.9。片段大小20–200 kb，平均长度50–150 kb（参见""）。DNA平均产量为150–500 µg，由血样中的核细胞数目决定。纯化的DNA可用于PCR、定量和real-time PCR（参见""）。

原理

PAXgene Blood DNA System是整合的标准化体系，用于收集和稳定全血样本并分离基因组DNA。该体系通过PAXgene Blood DNA Tubes收集和稳定血液，并使用PAXgene Blood DNA Kit纯化DNA。

PAXgene Blood DNA Tubes包含可防止血液凝结以及可稳定白细胞的混合试剂，用来收集血液。血液样本可在室温下在管中储存长达14天。用PAXgene Blood DNA Kit中提供的缓冲液获得的高纯度的基因组DNA适合各种下游应用。

程序

PAXgene Blood DNA操作步骤如流程图" PAXgene Blood DNA procedure"所示。血液样本（8.5 ml）由PAXgene Blood DNA Tubes收集，并储存或运输。如需纯化DNA，则将血液转入处理管中（已预充细胞裂解缓冲液），将溶液混合，裂解红细胞和白细胞。细胞核和线粒体通过离心收集、洗涤，重悬于裂解缓冲液中。蛋白污染物经蛋白酶孵育后去除。DNA用异丙酮沉淀、70%乙醇洗涤、干燥，并重悬于缓冲液。

应用

纯化的DNA可用于多种下游应用，包括：

PCR和定量real-time PCR
Southern印迹
SNP基因分型
药物基因组研究
SNP发现和鉴定

辅助数据和图表

PAXgene Blood DNA procedure.

Blood samples (8.5 ml) are collected in PAXgene Blood DNA Tubes, where they may be stored or transported. For DNA isolation, the blood is transferred to processing tubes (supplied already filled with cell lysis buffer), and the solution is mixed to lyse red and white blood cells. Cell nuclei and mitochondria are pelleted by centrifugation, washed, and resuspended in digestion buffer. Protein contaminants are removed by incubation with a protease. DNA is precipitated in isopropanol, washed in 70% ethanol, dried, and resuspended in resuspension buffer.

Efficient multiplex PCR of 3 mitochondrial genes.

High quality and high molecular weight of genomic DNA.

规格

特点	规格
Main sample type	Whole blood
Time per run	60 min/8 samples
Elution volume	1 ml
Yield	150–500 µg
Format	Processing tube
Sample amount	8.5 ml
Applications	(RT-)PCR, (RT-)qPCR, NGS, genotyping, microarray analysis
Processing	Manual (centrifugation)

资源

Second edition — innovative tools

For collection, storage, and transport of whole blood for genomic DNA isolation

Effect of preanalytical factors on analyte quality as shown with the QIAxcel Connect capillary gel electrophoresis system

Groelz et al., ASHG 2004

In order to reduce paper consumption and oblige the growing number of customers requesting an environmentally friendly alternative to traditionally printed handbooks, we are now providing kit handbooks for Research Use Only (RUO) PreAnalytiX kits on our website only.

This note is to inform you that the street address for PreAnalytiX GmbH has changed from “Feldbachstrasse” to “Garstligweg 8”. Please be informed that the update of the product labeling to the new address is ongoing.

Download Safety Data Sheets for QIAGEN product components.

常见问答

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ-12

The typical A260/A280 ratio is from 1.7 to 1.9. The DNA molecules are up to 200 kb in size, with fragments of 50–150 kb predominating.

FAQ-3497

Yes, mitochondrial and genomic DNA are both isolated.

FAQ-3498

No. DNA purified with the PAXgene Blood DNA System is free of detectable RNA contamination. In contrast to commercial salting out methods, additional RNase digestion steps are not necessary.

FAQ-3499

No. The volume of Buffer BG1 (lysis buffer) prefilled in the processing tubes, and all the other buffer volumes are optimized for use with blood from one PAXgene Blood DNA Tube.

FAQ-3500

Buffer BG4 consists of 10 mM Tris·Cl, pH 8.5.

FAQ-3501

DNA purified using the PAXgene Blood DNA System should be dissolved in Buffer BG4 and stored at 2–8°C. For long-term storage, DNA can be stored at –20°C but multiple freezing and thawing cycles should be avoided.

FAQ-3502

Yes. After dissolving the nuclei pellet in digestion buffer, Buffer BG3 (step 7), samples can be stored up to 7 days at 2–8°C or for one day at room temperature (18–25°C). The protocol can also be interrupted after adding isopropanol (step 10).

FAQ-3503

The system is designed for use only with human blood samples.

FAQ-3504

Do not add bleach or acidic solutions directly to the sample preparation waste. CAUTION: The sample preparation waste contains guanidinium hydrochloride from Buffer BG3, which can form highly reactive compounds when combined with bleach. For more information please consult the appropriate material safety data sheets (MSDS).

FAQ-3505

So far, we have shown that DNA purified using the PAXgene Blood DNA System can be used in PCR, including long range and multiplex PCR, restriction digestion and Southern analysis, as well as in various SNP detection procedures including,TaqMan®, LightCycler and Masscode technologies.

See trademarks.

FAQ-3506

See subsequent FAQs for information about storage interval and temperature. Each laboratory should validate shipping procedures.

FAQ-3507

Samples in PAXgene Blood DNA Tubes can be stored at room temperature (18–25°C) for up to 14 days after blood collection.

FAQ-3508

Yes, samples in PAXgene Blood DNA Tubes can be stored at 30°C for up to 3 days after blood collection. However, it is not recommended to store blood samples at temperatures higher than 30°C and storage at high temperatures should not exceed 3 days.

FAQ-3509

Yield of genomic DNA varies depending on the white blood cell count of the patient blood sample, the extraction method, and the elution volume. However, DNA yield typically ranges between 16–135 µg/ml whole blood. Specifically, DNA yields from magnetic bead-based extraction on the QIAsymphony ranges between 16.9–51.7 µg/ml whole blood, from silica membrane-based extraction on the QIAcube 17.1–45.8 µg/ml whole blood, and from salting-out precipitation 4.9–53.7 µg/ml whole blood.

FAQ-3510

The typical A₂₆₀/A₂₈₀ ratio is from 1.7 to 1.9.

FAQ-3511

To date, DNA purified from blood samples in PAXgene Blood DNA Tubes has been successfully tested for use in PCR, including multiplex PCR.

FAQ-3512

You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .

FAQ-3515

TB Management

Transplant

Infectious Disease

Oncology

Sexual & Reproductive Health

Sample Processing

Solutions for Laboratory-Developed Tests