New England Biolabs, T4070V, Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit

Related Categories Magnetic Bead-Based Nucleic Acid Purification,, Nucleic Acid Purification Applications Nucleic Acid Purification FAQ Q: What sample types can I use with the Monarch® Mag Cell-free DNA Extraction Kit (NEB #T4070)? A: This kit has been evaluated for plasma and urine samples. Other biological samples, such as cerebrospinal fluid (CSF), are also compatible. Q: Can I add Monarch® StabiLyse DNA/RNA Buffer (NEB #T2111) to plasma and store it for extraction later? A: Yes, Monarch StabiLyse DNA/RNA Buffer can be added to plasma in the proportions recommended in the Sample digestion step of the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070) manual, and the mixture stored at -20°C or -80°C until ready to proceed. Q: The Monarch® Mag Beads M2 (NEB #T4105) have settled down at the bottom of the bottle. Is this a problem? A: When undisturbed and not in use, it is normal for the Monarch Mag Beads M2 to settle down very quickly. This has no impact on the functionality of the beads. Before use, it is important to mix the beads thoroughly by vigorous vortexing to achieve homogeneous dispersion. Q: Can I purchase the components of the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4105) separately? A: Yes, all components of this kit are also available for purchase individually. Please see the kit components table with links to the respective product webpages or search for the component names on neb.com. Q: What is the composition of each reagent provided with the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070)? A: All reagent composition information is proprietary. Q: When using the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070), instead of adding Monarch Buffer BX, Isopropanol and the beads separately into the sample, can I make a binding mix of these three components? A: Yes, a binding mix of Monarch Buffer BX, Isopropanol, and Monarch Mag Beads M2 can be made for use within a day. Ensure it is well mixed before adding to the sample tube. Q: What is the lowest elution volume I can use when using the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070)? A: Optimal elution volume depends on the sample volume and the magnetic beads. For ideal recovery, the elution volume should be half of the volume of beads used although a lower elution volume, for example, one third of the bead volume used, also produces good recovery. Other factors to consider when choosing the elution buffer volume include the expected cfDNA content in the sample and the downstream application's input requirements. Q: When using the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070), what concentration of the DNA needed for downstream applications? A: Our recommended elution volumes typically yield sufficient output at the concentration needed for downstream NGS library preparation and help eliminate the need for an additional concentration step after cfDNA extraction. However, if a higher concentration is strongly desired, a protocol is available on the product webpage. Some yield loss may occur with additional concentration steps. Q: When using the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070), I noticed that wash buffer volumes are the same regardless of the sample input volume. Can I use lower volumes of wash buffers with low sample volume? 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For sample inputs less than 1 ml, reduced volume of the washes can be used if desired, for example, 500-700 µl. Q: I am interested in cfRNA extraction. Can I use the Monarch® Mag Cell-free DNA (cfDNA) Extraction Kit (NEB #T4070) for cfRNA extraction? A: The reagents and workflow provided with the Monarch Mag Cell-free (cfDNA) Extraction Kit are optimized for cell-free DNA extraction. Please direct inquiries about other applications, such as cfRNA extraction to NEB Technical Support. Q: I have used all the Proteinase K (NEB #P8200) included in the Monarch kit. Can I purchase more? A: Yes, Proteinase K, Molecular Biology Grade (NEB #P8107), is available for standalone purchase and can be used as a direct substitute for (NEB #P8200) in all Monarch protocols without any modifications.