QIAamp DSP DNA Blood Mini Kit

In vitro diagnostics를 목적으로한 human blood에서 genomic DNA을 분리 정제할 수 있습니다

특징

Universal DNA purification system compatible with other IVD products
여러 antidoagulants와 tube와 호환해서 사용할 수 있어서 sample collection 유동적으로 가능
Compatible with different anticoagulants and tubes
Dentrifuge protocol 또는 vacuum protocol중 선택 가능

제품 세부 정보

The QIAamp DSP DNA Blood Mini Kit provides silica-based DNA purification. The QIAamp DSP DNA Blood Mini Kit is designed for labs that process blood for in vitro diagnostic use.

성능

Genomic DNA purified using the QIAamp DSP DNA Blood Mini Kit is ready to use in downstream applications like those based on enzymatic amplification or other modification, such as PCR.

Whole blood samples can be collected using many types of anticoagulant-containing tubes (EDTA or citrate), including BD Vacutainer, Monovette, and Vacuette tubes (see table). Blood samples can be frozen and thawed at least 3 times and still be a reliable source for DNA purification (see figure " DNA from frozen and thawed blood").

Blood collection using a range of different tubes
Primary tube	Manufacturer	Cat. no.	Nominal volume	Average yield from 200 µl
BD Vacutainer 9NC	BD	366007	9 ml	6.4 µg
BD Vacutainer K3E	BD	368457	10 ml	6.6 µg
BD Vacutainer K2E	BD	367864	6 ml	6.4 µg
S-Monovette EDTA	Sarstedt	02.1066.001	9 ml	6.5 µg
S-Monovette CPDA1	Sarstedt	01.1610.001	8.5 ml	6.3 µg
Vacuette K3E	Greiner Bio-One	455036	9 ml	6.5 µg
Vacuette 9NC	Greiner Bio-One	454382	2 ml	6.3 µg

그림 참조

DNA from frozen and thawed blood.

원리

The QIAamp DSP DNA Blood Mini Kit uses well-established QIAamp technology for purifying genomic DNA. The QIAamp silica-based membrane specifically binds DNA in the lysed sample, while the rest of the lysate is rapidly removed by either centrifugation or vacuum. The bound DNA is efficiently washed to remove contaminants and then eluted in volumes of 50–200 µl.

절차

Genomic DNA is purified from blood using one of 2 alternative procedures, using a centrifuge or using a vacuum manifold and centrifuge (see flowchart " Procedure"). The universal DNA purification system allows compatibility with other in vitro diagnostic products (see " Workflow").

The EDTA- or citrate-containing blood sample (200 µl) is lysed in the presence of QIAGEN Protease and lysis buffer at 56°C for 10 minutes. Ethanol is then added to the lysate to optimize binding of DNA to the QIAamp membrane. The lysate is applied to a QIAamp Mini spin column, which is then centrifuged or subjected to vacuum pressure. DNA binds to the QIAamp membrane in the spin column, and the rest of the lysate passes through. Bound DNA is efficiently washed by 2 different wash buffers, which are also drawn through the QIAamp membrane by centrifugation or vacuum pressure. The QIAamp membrane is then dried by centrifugation. Elution buffer (50–200 µl) is applied to the QIAamp membrane and, after a 1-minute incubation, the spin column is centrifuged to elute pure DNA.

Purification of DNA using the QIAamp DSP DNA Blood Mini Kit can be fully automated on the QIAcube. If automating the QIAamp DSP DNA Blood Mini Kit on the QIAcube instrument, the instrument may process fewer than 50 samples due to dead volumes, evaporation, and additional reagent consumption by automated pipetting. QIAGEN only guarantees 50 sample preps with manual use of the QIAamp DSP DNA Blood Mini Kit.

그림 참조

QIAamp DSP DNA Blood Mini Kit procedure.

QIAamp DSP DNA Blood Mini Kit workflow.

응용 분야

The QIAamp DSP DNA Blood Mini Kit provides proven QIAamp technology for purification of DNA from fresh or frozen whole blood, as well as blood that has been treated with citrate or EDTA.

지원되는 데이터 및 수치

DNA from frozen and thawed blood.

EDTA-treated blood samples were frozen and thawed up to 3 times. DNA was purified at the end of each freeze-thaw cycle. DNA was purified from 200 µl samples using the QIAamp DSP DNA Blood Mini Kit and eluted in 200 µl elution buffer. DNA yields are normalized to the yield from fresh sample (100%). Each bar represents the results from 32 replicates (mean ± standard deviation).

Specifications

Features	Specifications
Applications	Downstream detection procedures in molecular diagnostics, such as PCR
Main sample type	Whole blood
Elution volume	50–200 µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Genomic DNA
CE/FDA/IVD compatible	CE/IVD
Format	Spin columns
Sample amount	200 µl
Processing	Manual (centrifugation or vacuum); automated (on QIAcube Connect MDx)
Technology	Silica technology

리소스

FAQ

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

FAQ ID -761

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728

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 ID -12