QIAamp MinElute ccfDNA Kits

ヒト血漿あるいは血清から遊離DNAを濃縮・精製

S_1084_5_GEN_V2

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

QIAamp MinElute ccfDNA Midi Kit (50)

Cat. No. / ID:  55284

For 50 preps (4 or 5 ml sample input volume each): QIAamp UCP MinElute Columns, QIAGEN Proteinase K, Magnetic Bead Suspension, Buffers, Bead Elution Tubes, and Collection Tubes (1.5 ml and 2 ml)
Copy order details
$1,312.00
Log in To see your account pricing.
KitAccessories
QIAamp MinElute ccfDNA Kit
Accessories
Volume
5–10 ml
1–4 ml
日本では本採血管は医療機器となります。販売に関しては 日本ベクトン・ディッキンソン株式会社へお問い合わせください。
QIAamp MinElute ccfDNA Kitsは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 断片化DNA を効率的に回収
  • サンプルインプット量を増やし(最大10 ml)、溶出量を減らす(最少20 μl)ことで核酸の高濃縮を 実現
  • 高収量で確実に低存在量のバイオマーカーを検出
  • 有機溶媒抽出およびアルコール沈殿不要、夾雑物および阻害物質を効率よく除去

Product Details


QIAamp MinElute ccfDNA Kits は、血清/血漿中の 遊離DNA(ccfDNA; circulating cell-free DNA)を簡単に濃縮・精製できます。精製ステップは QIAcube による自動化が可能です。

Performance

血漿、血清および体液中のccfDNA の存在量は大変に低く、通常1,000 bp 未満の短い断片として存在しています(ヒト血漿中 ccfDNA 1 ~ 100 ng/ml)。そのため単離は非常に困難です。QIAamp MinElute ccfDNA Kits は、少量であっても、血清および血漿サンプルから高品質のccfDNAを効率的に精製し、優れた結果を提供します。他のキットと比較して、QIAamp MinElute ccfDNA Kits は、最大のccfDNA 収量を示しました(図  最も高いccfDNA 収量)。遊離DNA(ccfDNA)を磁性ビース法により、幅広い容量のサンプル(1 ~10 ml)から高度な濃縮が可能です(図  様々なスタートサンプル量に対応可能)。また、精製ステップは定評のあるカラムメンブレン(QIAamp UCP MinElute Columns)で、溶出量も20〜80 µl まで様々に対応できます。QIAamp MinElute ccfDNA Mini Kitは1〜4 ml、QIAamp MinElute ccfDNA Midi Kitは4〜10 ml のサンプル量に至適化されています。QIAamp UCP MinElute Column は20塩基程度の短い断片化DNAを結合する事が出来ますが、収量についてはサンプル中のccfDNA の濃度とサンプルの状態に依存します。
See figures

Principle

血液中の血中循環腫瘍DNA(ctDNA)や、母体血液中の胎児DNAのようなccfDNA は、通常1,000 bp 未満の短い断片として血清または血漿中に存在しています(ヒト血漿中 ccfDNA 1 ~ 100 ng/ml)。特にctDNA は腫瘍ゲノム全体の情報をもち、有効な臨床応用の可能性があります。血漿、血清などの体液中のccfDNA 濃度は、大変低く、その濃縮、精製には依然として大きな課題があります。QIAamp MinElute ccfDNA Kits は、QIAamp Circulating Nucleic Acid Kit の性能を継承しながら、磁性ビーズ方式で簡便にccfDNA 濃縮を行ない、大量サンプル量(1~10 ml)への適応を可能にしました。ccfDNA 精製のステップは QIAcubeで自動化することができます。

Procedure

本キットは迅速かつ簡便な磁性ビーズ方式によるccfDNA 濃縮ステップにより、サンプル間のクロスコンタミネーションを排除し、潜在的に感染性のあるサンプルを取り扱う際の安全性を高めます。簡便な操作で、複数サンプルの同時処理に適しており、24 サンプルを1 時間未満で精製できます。QIAamp MinElute ccfDNA Mini Kit は2 ml のサンプル処理に至適化されたプロトコールを採用しており、4 ml のサンプルを処理にする場合には1 キットあたりのサンプル処理数は25 回となります。 同様にQIAamp MinElute ccfDNA Midi Kit は4 ml のサンプル処理に至適化され、10 ml のサンプルを処理する場合には、1 キットあたりのサンプル処理数は25 回分となります。
Ultra-Clean Water で溶出された溶出液は、タンパク質などの不純物を含まない核酸抽出液で、–15 ~ –30℃での即時保存 も可能です。
本キットは、血清および血漿サンプルから最大のccfDNA 収量、高濃度、最短時間で、高品質ccfDNA を得られていることを示しています(図  迅速なccfDNA精製 最も高いccfDNA収量 高濃度なccfDNA)。
 

精製ステップはQIAcubeによる自動化が可能です。
See figures

Applications

QIAamp MinElute ccfDNA Kits は、ヒト血清/血漿中ccfDNA の効率的な精製に至適な製品です。

Supporting data and figures

Resources

MSDS (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
キットハンドブック (1)
クイックスタートプロトコール (2)
Certificates of Analysis (1)
Kit Handbooks (1)

FAQ

Which protocol line should I use to process plasma samples?
The QIAsymphony PAXgene Blood ccfDNA Kit can be sued with 2 protocol lines. The standard protocols (STA) predominantely isolate small ccfDNA fragments. The large fragment protocols (LAF) efficiently co-purify large and small ccfDNA fragments. Both lines include a protocol for 2.4 ml and another for 4.8 ml plasma input.
FAQ ID - 3654
Which elution volumes are available with the QIAsymphony PAXgene Blood ccfDNA Kit?
The elution volume of the standard protocols (STA) is fixed at 60 µl. The large fragment protocols (LAF) enable elution in 3 different volumes: 60, 100 and 150 µl.
FAQ ID - 3655
What is the shelf life of the QIAsymphony PAXgene Blood ccfDNA Kit?
The expiration date is printed on the kit label and encoded in the bar code of the reagent cartridges.
FAQ ID - 3651
What is the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) and what is the impact for me as a customer?
For overview and further information on the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) please visit www.qiagen.com/ghs .
FAQ ID - 3428
Can whole blood in PAXgene Blood ccfDNA Tubes be stored at temperatures above 25°C?
Whole blood drawn into PAXgene Blood ccfDNA Tubes can be stored for at least 24 hours at 35°C. Higher temperatures will lead to lysis of blood cells and release of genomic DNA into plasma.
FAQ ID - 3640
Which downstream applications have been tested with ccfDNA purified from whole blood collected and processed with the PAXgene Blood ccfDNA System?
To date, ccfDNA from blood samples collected and processed using the PAXgene Blood ccfDNA System have been successfully tested in PCR, including multiplex qPCR, and NGS applications.
FAQ ID - 3658
Which tubes are recommended for the QIAsymphony sample rack
The recommended sample input format is the 14 ml Falcon® polystyrene round bottom tube 17 x 100 mm supplied by Corning (formerly supplied by BD).
FAQ ID - 3653
Can whole blood be frozen in PAXgene Blood ccfDNA Tubes?
No. Freezing blood drawn into PAXgene Blood ccfDNA Tubes results in lysis of red and white blood cells and release of genomic DNA.
FAQ ID - 3642
Are PAXgene Blood ccfDNA Tubes sterile?
Yes, the inside of PAXgene Blood ccfDNA Tubes is sterilized during production.
FAQ ID - 3636
What is the shelf life of unused PAXgene Blood ccfDNA Tubes?
The expiration date is printed on the tube label and on the shelf carton. 
FAQ ID - 3633
How long after first use can a reagent cartridge be stored for further use?
When closed properly with the provided strips and stored at room temperature (15–25°C), a reagent cartridge can be used up to 6 weeks after the initial run.
FAQ ID - 3650
Can whole blood in PAXgene Blood ccfDNA Tubes be stored at temperatures below 15°C?
Yes. Whole blood in PAXgene Blood ccfDNA Tubes can be stored at 8–15°C for up to 7 days. Storing blood in PAXgene Blood ccfDNA Tubes below room temperature does not extend stability. 
FAQ ID - 3641
What should I do if cryoprecipitates form while thawing a frozen plasma sample?

If cryoprecipitates cannot be resolved completely by vortexing, the sample should be centrifuged to prevent precipitates from blocking the column. To ensure that no cfDNA is co-precipitated,  centrifugation can be performed at a lower speed (at 6,800xg for 3 minutes at room temperature, as stated in the Protocol Sheet for the QS DSP Circulating DNA Kit). 



FAQ ID - 3646
How long and at which temperature can plasma generated from whole blood in PAXgene Blood ccfDNA Tubes be stored?
Plasma generated from whole blood collected into PAXgene Blood ccfDNA Tubes can be stored at 28°C for up to 14 days. We recommend freezing plasma at 20°C or 80°C for longer storage periods (long-term storage studies are ongoing).
FAQ ID - 3644
What is the expected ccfDNA yield from plasma generated from 10 ml of blood?
The yield of ccfDNA depends on the target ccfDNA. Nevertheless, yields obtained from PAXgene Blood ccfDNA Tubes are comparable to those obtained from the same volume of plasma separated directly after drawing blood into a spray-dried EDTA tube. In the PAXgene Blood ccfDNA Tube, however, this yield remains stable for up to 7 days when the tube is stored at 15–25°C or for up to 24 hours when stored at 35°C. In contrast, genomic DNA is released from lysed and apoptotic cells in the unstabilized contents of the EDTA tube soon after blood draw.


FAQ ID - 3637
Do I need to use a blood collection set with the PAXgene Blood ccfDNA Tube?
Yes. Further information on phlebotomy technique with the PAXgene Blood ccfDNA Tube can be found in the PAXgene Blood ccfDNA Tube Product Circular under the Resources tab.
FAQ ID - 3626
How many extractions can I perform with a QIAsymphony PAXgene Blood ccfDNA Kit, using the 2.4 ml or the 4.8 ml protocols?
The number of extractions that can be performed with a kit is independent of sample volume. A kit contains sufficient reagents for 192 extractions.
FAQ ID - 3649
Can reagent from the PAXgene Blood ccfDNA Tube flow back into the patient’s arm?
If the instructions for use are followed, there is no possibility of backflow during phlebotomy and thus, no possibility of reagent going into the patient’s arm.
FAQ ID - 3629
Can I use plasma from underfilled PAXgene Blood ccfDNA Tubes?
A complete 10 ml blood draw ensures optimal results. However, depending on the downstream application and target ccfDNA fragment size, tubes filled with as little as 6.5 ml whole blood can be used without adversely affecting results.
FAQ ID - 3638
Can I use a different brand blood collection set with the PAXgene Blood ccfDNA Tube?
Yes, as long as the phlebotomy instructions included with the PAXgene Blood ccfDNA Tubes are properly followed.
FAQ ID - 3630
What is the reagent in the PAXgene Blood ccfDNA Tube?
The PAXgene Blood ccfDNA Tube contains a proprietary formulation that prevents blood coagulation and the release of genomic DNA from blood cells into plasma. Contents of the tube may cause irritation to the eyes, respiratory system and skin. Please refer to the PAXgene Blood ccfDNA Tube Product Circular. For additional information, visit www.preanalytix.com or review the Safety Data Sheet under the Resources tab.
FAQ ID - 3632
What is the blood draw volume of the PAXgene Blood ccfDNA Tube
The blood draw volume is 10 ml and the stabilization reagent volume is 1.5 ml. Thus, a filled tube holds a volume of 11.5 ml.
FAQ ID - 3631
For what type of sample material is the QIAsymphony PAXgene Blood ccfDNA Kit intended?
The QIAsymphony PAXgene Blood ccfDNA Kit is designed to be used with plasma generated from whole blood collected into PAXgene Blood ccfDNA Tubes.
FAQ ID - 3652
How should I thaw frozen plasma samples?
Thaw the tubes at room temperature (15–25°C). To minimize the formation of cryoprecipitates, do not thaw at lower temperatures (e.g., 4°C). Alternatively, thaw plasma for 30 minutes at 30°C to completely avoid the formation of cryoprecipitates.
FAQ ID - 3645
Are there special considerations for phlebotomy when using the PAXgene Blood ccfDNA Tube (blood collection set, discard tube, positioning of the tube, issues with short draws, etc.)?

Yes. Following a standard phlebotomy protocol, blood is collected into the evacuated tube using a blood collection set and a standardized needle holder that can accommodate a 16 mm diameter tube. Make sure that blood has stopped flowing into the tube before removing the tube from the holder.

For more information on phlebotomy technique, please refer to the PAXgene Blood ccfDNA Tube Product Circular under the Resources tab.
FAQ ID - 3635
Is double centrifugation required for plasma processing?
No. However, a second centrifugation step helps remove traces of buffy coat that may be accidentally transferred when aspirating plasma after the first centrifugation step.
FAQ ID - 3643
How should eluates produced with the PAXgene Blood ccfDNA System be stored?
Eluates should be stored at 20°C (long-term storage studies are ongoing).
FAQ ID - 3657
Does the QIAsymphony PAXgene Blood ccfDNA Kit also allow eluting ccfDNA in less than 60 µl?
Yes, a customized protocol for elution with 50 µl is available on request.
FAQ ID - 3656
What type of magnet do you recommend for handling the magnetic beads in the Qproteome Plasma Membrane Protein protocol?

Since the Qproteome Plasma Membrane Protein procedure requires 1.5 ml/2.0 ml reaction tubes, we recommend using the follwing magnets for handling the magnetic beads:

Alternatively, comparable tube magnets or a standard clip-board magnet should also work.

FAQ ID -1193
How long can whole blood stabilized in PAXgene Blood ccfDNA Tubes be stored at room temperature?
Whole blood drawn into PAXgene Blood ccfDNA Tubes can be stored at 15–25°C for up to 7 days.
FAQ ID - 3639
Why have the hazard symbols accompanying products changed?
The Globally Harmonized System (GHS) is an initiative by the United Nations aiming to harmonize chemical hazard communication worldwide. Hazards symbols accompanying products are changing to those of the GHS. Information on the adoption of this new system is available on our GHS Hazard Labeling pages.
FAQ ID - 3430
Which plasma volumes can I process with the QIAsymphony PAXgene Blood ccfDNA protocols?
The normal protocols enable processing 2.4 or 4.8 ml plasma. Other volumes can be accommodated using customized protocols. 
FAQ ID - 3648
How should unused PAXgene Blood ccfDNA Tubes be stored?
Store PAXgene Blood ccfDNA Tubes at room temperature (15–25°C) and protect them from direct sunlight.
FAQ ID - 3634
Why is a blood collection set required to collect blood into the PAXgene Blood ccfDNA Tube?
When used in accordance with instructions, the blood collection set eliminates the possibility of patient contact with the reagent in the tube due to backflow of blood.
FAQ ID - 3628