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View additional product information for Sequential Protein/DNA/RNA Extraction Kit - FAQs (50000D, 50010D)
24 product FAQs found
The lysis buffer included in the kit has been specifically optimized for efficient lysis of cells and tissue samples and should not be replaced with another lysis buffer. However, it can be supplemented with protease inhibitors. RNase inhibitors are not necessary, as RNA integrity is well maintained.
The protein, DNA and RNA eluates exhibit high purity with very low carry-over between fractions.
Yes, the kit can be used with a plethora of sample types, provided the sample is sufficiently lysed.
There is no available protocol for on-bead digestion, although the kit is theoretically compatible with this process. The kit can be combined with the EasyPep MS Sample Prep Kit by skipping the lysis step.
Dynabeads Sequential Protein Binding Beads have a diameter of 2.8 µm, while MagMAX Sequential DNA/RNA Binding Beads have a diameter of 1 µm. The binding capacity cannot be disclosed here, as it involves proprietary information regarding the binding principle.
Yes, the remaining lysate from the sequential workflow can be stored at -80°C and used later without impacting the yield and quality of the analytes.
Yes, the yields obtained using the kit are comparable to those obtained with single analyte isolation kits. However, the yields of proteins are usually slightly lower than cells lysed directly with RIPA buffer.
The buffers and protocols have been optimized to help ensure good RNA integrity.
The kit enables high sensitivity from small sample input sizes, making it exceptional for precious samples (e.g., patient samples) or samples that cannot be split for separate extraction of protein, DNA, and RNA (e.g., heterogeneous tissue specimens).
The manual protocol uses ~2 hours of hands-on time for 10 samples. The automated protocol uses ~1.5 hours of instrument run time and 30–60 minutes of hands-on time for 12 to 96 samples.
The kit is magnetic bead-based and thus compatible with liquid handlers (e.g., Hamilton NIMBUS Presto System).
The kit uses magnetic beads for the specific isolation of protein, DNA, and RNA, which enables automation of the workflow, resulting in higher reproducibility and throughput. Additionally, our optimized buffer chemistry increases sensitivity, allowing the use ofsmaller sample sizes from the start. Lastly, the protein can be directly prepared for mass spectrometry analysis without further desalting.
The kit is developed for cells and is not directly compatible with whole blood samples. We recommend pre-isolating cells from blood, such as peripheral blood mononuclear cells (PBMCs), prior to protein, DNA, and RNA extraction for optimal results.
While these sample types have not been specifically tested with our kit, theoretically they should work, provided the sample is sufficiently lysed. Proper lysis is crucial to help ensure effective isolation of protein, DNA, and RNA from fungi, plant cells, or yeast.
The kit is compatible with targeted sequencing (e.g., Ion Torrent next-generation-sequencing (NGS) systems and short-read sequencing (e.g., Illumina NGS platforms).
The kit has not been assessed for long-read sequencing.
The kit is not compatible with the Olink PEA technology, since this requires native proteins for capture by a pair of antibodies, whereas our kit denatures the proteins for mass spectrometry analysis.
The kit has not been optimized for use with FFPE samples. Additionally, FFPE samples are not ideal for RNA integrity, which may affect the quality and yield of RNA isolated from these samples. For optimal results, we recommend using fresh or frozen tissue samples.
Yes, this kit is designed to isolate total RNA, which includes small RNAs like microRNAs.
The size of the isolated proteins ranges from around 5 to 590 kDa. The isolation of specific peptides has not been assessed.
Yes, the lysis buffer’s effectiveness has been tested by diluting it up to 2 times, with performance like the non-diluted lysis buffer.
The kit is optimized for a maximum input of 1,000,000 cells or 10 mg of tissue per extraction.
The kit is optimized for 100,000 cells and 1 mg of tissue. We have tested down to 1,000 cells and 0.005 mg of tissue, demonstrating linearity in the yields of protein, DNA, and RNA.
Yes, the sequential workflow can be paused after protein isolation and after DNA isolation by freezing the sample plate at -20°C, without impacting the yield and quality of the analytes.