Sequentially isolate DNA and RNA from blood and bone marrow

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Hematologic malignancies account for 9.4% of new cancer cases each year [1]. Isolation of total nucleic acids from hematological samples is crucial in understanding and identifying cancer causing genetic alterations.

A comprehensive genomic profiling approach using both DNA and RNA is necessary to detect the various types of genetic abnormalities found in hematologic malignancies. The Applied Biosystems MagMAX Sequential DNA/RNA kit utilizes the power of Dynabeads magnetic bead technology to enable the sequential purification of high-quality genomic DNA and total RNA from a single whole blood or bone marrow sample for your cancer research.

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What is hematologic cancer research?

Hematological cancer research seeks to enhance our understanding of the underlying genetic, molecular, and cellular mechanisms involved in these diseases. By utilizing innovative research methods and therapeutic approaches, researchers aim to improve outcomes, develop more effective early detection strategies, and ultimately discover cures for these complex and challenging malignancies. The three most common blood disorders are Anemia, thrombocytopenia and leukemia, whereas hemophilia is an example of a rare blood disorder.

Learn more about hematological cancer clinical research

Synergetic biomarkers: Fueling advancements in cancer research

Oncology biomarker applications have helped transform our understanding and approach in the study of cancer. Explore this evolving field through the lens of these biomarkers and the advancements enabling breakthrough discoveries.

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Advantages of sequentially isolating nucleic acid over isolation of single analyte or total nucleic acid

Molecular profiling, which helps provide a comprehensive profile at the molecular level, begins with isolation of both DNA and RNA. Total nucleic acid (TNA) isolation from hematological samples can help offer insights into genetic alteration relevant to blood cancers. Isolating these analytes sequentially helps researchers to:

  • Isolate separate DNA and RNA eluates—eliminates the need for additional steps, including manual RNase treatment to a final TNA eluate
  • Increase productivity—automation of sequential DNA and RNA isolation provides a streamlined workflow for faster time-to-date with less hands-on time
  • Make workflows scalable—enables high-quality nucleic acid isolation, offering scalable and flexible sample volume input and throughput
  • Optimize test utilization management—when triaging a low volume sample, previous blood or bone marrow samples can be saved by utilizing a sequential workflow


MagMAX Sequential DNA/RNA Kit

The MagMAX Sequential DNA/RNA Kit helps maximize DNA and RNA yields from precious samples with separate, ready-to-use elutes for a broad range of molecular applications including qPCR and sequencing. The kit is compatible with KingFisher sample purification systems for a scalable workflow to enable fast time-to-date while meeting specific throughput needs.

A complete kit or standalone components are available for larger volume processing.


Sequential workflow

The MagMAX Sequential workflow isolates DNA and RNA sequentially from whole blood or bone marrow sample types on KingFisher Duo Prime, KingFisher Flex or KingFisher Apex purification systems. The total turnaround time is ~2.5 hours with total hands-on time* of ~45 minutes.

*Hands-on time may vary due to sample number and user method for sample addition to the sample plate.

  • Step 1: Set up the KingFisher sample purification system and prepare your whole blood or bone marrow samples and on-board digestion with proteinase K.
  • Step 2: Prepare DNA lysis/binding and bead mix. Add it to each sample.
  • Step 3: Continue DNA extraction protocol on KingFisher Sample Purification System. Obtain purified DNA.
  • Step 4: Now, to obtain purified RNA, prepare RNA binding and bead mix. Add it to RNA-containing supernatant (DNA sample plate).
  • Step 5: Set up KingFisher sample purification system and load plates on it.
  • Step 6: After on-board DNAase step, add RNA re-binding solution. Obtain purified RNA.

Figure 1. Sequential DNA and RNA isolation from whole blood or bone marrow using MagMAX Sequential DNA/RNA kit. Automate the workflow using either KingFisher Duo Prime, KingFisher Flex or KingFisher Apex Purification Systems.

Complete the workflow using Oncomine Myeloid Research Assay, Ion Chef System and Ion Genestudio S5 system.

Figure 2. Purified eluates are compatible for downstream analysis using Oncomine Myeloid Research Assay, Ion Chef System and Ion Genestudio S5 system. Once purified, the nucleic acid library can be prepared using the Oncomine Myeloid Research Assay and the template can be prepared on the Ion Chef System. Once completed, conduct next-generation sequencing (NGS) on the Ion GeneStudio S5 system and analysis using a reporter software. By integrating the MagMAX Sequential DNA/RNA Kit with KingFisher Sample Purification Systems with the downstream solutions, you can obtain a comprehensive view of the DNA mutations and gene fusions for myeloid malignancies.



Performance data

Sequential isolation performance data for DNA and RNA from bone marrow and whole blood sample types using the MagMAX Sequential DNA/RNA Kit

Bone Marrow

graphed results

Whole Blood

graphed results

Figure 3. DNA averaged across multiple samples across various sample volume input. DNA measured on Invitrogen Qubit Flex Fluorometer from Invitrogen Qubit 1X dsDNA Broad Range Assay. Achieve >200 ng DNA with 100 µL whole blood or 50 µL bone marrow while obtaining high-yielding, high-quality DNA with scalable inputs. Note that scalability does not reflect linearity and donor to donor variation is expected and dependent on sample quality, age, and input.

Bone Marrow

graphed results

Whole Blood

graphed results

Figure 4. RNA averaged across multiple samples across various sample volume input. DNA measured on Invitrogen Qubit Flex Fluorometer with Invitrogen Qubit RNA High Sensitivity (HS) Assay. Please note that RNA yields are variable from donor to donor and dependent on sample quality, age, and input. Achieve >100 ng RNA from 100 µL whole blood or 50 µL bone marrow while obtaining high-yielding, high-quality RNA with scalable inputs. Note that scalability does not reflect linearity and donor to donor variation is expected and dependent on sample quality, age, and input.



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References

For Research Use Only. Not for use in diagnostic procedures.

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