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View additional product information for MagMAX™ FFPE DNA/RNA Ultra Kit - FAQs (A31881)
23 product FAQs found
许多因素都可能影响从FFPE组织提取的DNA的总体质量和得率。以下为我们就如何解决这些关键影响因素而提出的一些建议：
1.组织获取和样本制备 - 在条件允许的情况下，组织应在外科切除下来后一小时内固定。使用中性福尔马林或多聚甲醛缓冲液固定的最佳时长为12-24小时。固定后的组织进入包埋程序之前，应彻底脱水。
2.组织块的储存 -在条件允许的情况下，储存的组织块尽量保证无切面。如果出现切面，氧气、水、光、以及外源感染（如真菌、昆虫等）等其他环境因素可能会造成组织块的进一步破坏。
3.用于DNA分离的组织类型、大小和数量 – 建议的组织厚度为10-20 µm。所用的切片数量取决于组织类型（决定细胞密度）和表面积大小(推荐大小：50-300 mm2）。过量的起始材料可能导致过滤柱堵塞，导致得率下降。
4.包埋组织时的过量石蜡 - 在条件允许的情况下，在纯化操作之前脱去组织样本上过多的石蜡。对于二甲苯纯化法而言，室温条件下的两次二甲苯处理通常能够彻底脱石蜡。必要情况下，可在更为强效的37–55°C条件下处理达30分钟。在二甲苯脱蜡操作完成后，须对沉淀物进行两次100%乙醇漂洗，再彻底去除100%乙醇。磁珠法采用全新的化学方法来处理石蜡，所选用的组织切片应在20 µm以内。
请点击此处（https://www.thermofisher.com/us/en/home/references/ambion-tech-support/rna-isolation/general-articles/extraction-of-nucleic-acids-from-ffpe-samples.html）阅读更多关于FFPE样本核酸提取的信息。
我们为此提供了两种试剂盒：用于FFPE组织的RecoverAll总核酸分离试剂盒和MagMAX FFPE DNA/RNA Ultra试剂盒。请点击此处（http://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/dna-extraction/genomic-dna-extraction/dna-extractions-working-with-ffpe-samples.html）了解更多关于这些试剂盒的差异。
There are a number of factors that can impact the overall quality and yield of DNA isolated from FFPE tissues. Here are recommendations to address several key factors:
- Upstream tissue procurement and tissue specimen preparation - if possible, tissues should be fixed within one hour of surgical resection. The optimal fixation time is 12-24 hours using neutral-buffered formalin or paraformaldehyde. Fixed tissues should be thoroughly dehydrated prior to the embedding process.
- Block storage - storage of blocks without cut faces, when possible, prevents ongoing damage from exposure to atmospheric oxygen, water, and other environmental factors such as light and infestation (fungi, insects, etc.).
- Tissue type, size, and amount being used for DNA isolation - the recommended tissue thickness is 10-20 µm. The number of sections used is determined by the tissue type (which impacts cell density) and surface area (recommended size: 50-300 mm^2). Excess starting material can cause filter clogging, resulting in poor yield.
- Excessive amount of paraffin used for embedding tissues - when possible, excess paraffin should be trimmed away prior to starting the purification protocol. For xylene-based purification methods, two xylene treatments at room temperature should be sufficient for complete deparaffinization. If desired, a more rigorous 37-55 degrees C treatment can be performed for up to 30 minutes. After the xylene deparaffinization, it is crucial that the 100% ethanol is completely removed and the pellets are dry after the two 100% ethanol washes. The magnetic bead method employs novel chemistries to deal with the paraffin that limits input to 20 µm sections.
Read more about extraction of nucleic acids from FFPE samples here (http://www.thermofisher.com/us/en/home/references/Invitrogen-tech-support/rna-isolation/general-articles/extraction-of-nucleic-acids-from-ffpe-samples.html).
We offer 2 kits: RecoverAll Total Nucleic Acid Isolation Kit for FFPE and MagMAX FFPE DNA/RNA Ultra Kit Read more about the differences between these kits here (http://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/dna-extraction/genomic-dna-extraction/dna-extractions-working-with-ffpe-samples.html).
There is no specific upper limit to curl size for use with the MagMAX FFPE DNA/RNA Ultra kit (Cat. No. A31881). If all of the tissue is able to be submerged in the 200 µL of buffer, then the large volume of protocol should work. Please keep in mind when processing sections that are thicker than 40 µm, additional digestion buffer, binding solution, and wash buffer, may need to be purchased separately (Cat. No. A32796).
No. The AutoLys M tubes don't fit in a standard 1.5 mL or 2.0 mL heat block. We recommend incubating the tubes in the tube racks in an incubator or oven.
Any centrifuge with plate adapters will work. The AutoLys M Tube Racks fit nicely into those adapters.
No, the AutoLys M Tubes can be lifted manually, but the tools significantly reduce hand strain and increase ease of use as well as throughput.
No, they have to be purchased separately together with some other products that are necessary for using the AutoLys M Tube system efficiently:
- AutoLys M Tubes and Caps (Cat. No. A38738)
- Autolys M Tube Locking Lid (Cat. No. A37954)
- AutoLys M TubeLifter (Cat. No. A37956)
- AutoLys M Tube Pliers (Cat. No. A38261)
- AutoLys M Tube Racks (Cat. No. A37955)
The FFPE-associated wax and debris are held in the upper chamber of the AutoLys M tube while the lysate passes through. Nucleic acids can then be purified from the clarified lysate. The AutoLys M tubes thus reduce manipulation of the sample eliminating the need to pellet the tissue and wash, so tissue loss is minimized
The AutoLys M Tube system has only been tested with the MagMAX FFPE DNA/RNA Ultra Kit, but it may also work with our RecoverAll Total Nucleic Acid Isolation Kit for FFPE .
Yes, we offer the AutoLys M Tube system, which creates cleared lysates from FFPE tissue samples without the need for deparaffinization or wash steps, while increasing DNA and RNA yields.
We recommend the following DNA isolation kits:
- RecoverAll Total Nucleic Acid Isolation Kit for FFPE (Cat. No. AM1975)
- Ion Ampliseq Direct FFPE DNA Kit (Cat. Nos. A31133, A31136)
- MagMAX FFPE DNA/RNA Ultra Kit (Cat. No. A31881)
- PureLink Genomic DNA Mini Kit (Cat. Nos, K182000, K182001, K182002)
We highly recommend using the elution buffer that is included in the kit. Our R&D team has tested nuclease-free water for elution and has found it not to be as efficient resulting in more bead clumping and lower RNA/DNA yield.
Yes, we have a protocol for isolating DNA only (http://tools.thermofisher.com/content/sfs/manuals/MAN0015905_MagMAX_FFPE_DNA_only_Ultra_UG.pdf) and RNA only (http://tools.thermofisher.com/content/sfs/manuals/MAN0015906_MagMAX_FFPE_RNA_only_Ultra_UG.pdf).
Yes, we have a protocol (https://tools.thermofisher.com/content/sfs/manuals/MAN0015877_MagMAX_FFPE_DNA_RNA_Ultra_UG.pdf) for sequentially isolating DNA and RNA from the same FFPE sample.
Protease Digestion Buffer, Binding Solution, and DNA Wash Buffer are also available as standalone Cat. No. A32796. For processing sections that are thicker than 40 µm, additional reagents would be required.
Yes. We recommend using sections of 10 µm or thicker for miRNA extraction.
No, the buffers of the two kits are quite different and cannot be used interchangeably.
We recommend using the MagMAX FFPE DNA/RNA Ultra Kit since it is the most versatile and optimized kit. In addition to just isolating RNA or DNA, it allows for sequential isolation of RNA and DNA from the same FFPE sample.
The MagMAX FFPE DNA/RNA Ultra Kit allows for the sequential isolation of RNA and DNA from the same FFPE sample. The MagMAX FFPE Total Nucleic AcidIaolation kit requires you to perform separate protocols for RNA and DNA extraction using separate FFPE sections.
RNAse digestion is not needed because the DNA binding step has been optimized so that minimal RNA is pulled into the DNA fraction. For most downstream applications, some residual RNA should not cause any interference.
Yes, xylene can be used as well. Citrisolv Clearing Agent is a healthier alternative to xylene as it does not need to be handled under a fume hood.