New England Biolabs, T1120L, Monarch® Spin DNA Gel Extraction Kit

Related Categories DNA Gel Extraction,, Nucleic Acid Purification Applications Nucleic Acid Purification FAQ Q: Can I purchase additional Monarch® columns and collection tubes for the MonarchSpin DNA Gel Extraction Kit (NEB #T1120) and the Monarch Spin PCR & DNA Cleanup Kit (NEB #T1130) separately? A: Yes, columns and collection tubes that are compatible with DNA gel extraction and PCR cleanup (NEB #T2037L) are also sold separately. Q: Can I use water to elute the DNA when using Monarch® Kits? A: Yes, water can be used to elute DNA from Monarch columns. For maximum elution efficiency, ensure the water is nuclease-free. If you are storing the DNA long-term, we recommend using the supplied Monarch Buffer EY (DNA elution buffer), containing 0.1 mM EDTA. Q: What is the smallest volume of elution buffer that can be used with the Monarch® Spin Columns S1A (NEB #T2037)? A: The optimal range for elution volume is 5 - 20 μl of elution buffer. Q: Are the columns in the Monarch® Spin PCR & DNA Cleanup Kit (5 μg) (NEB #T1130) the same as those in the Monarch Spin DNA Gel Extraction Kit (NEB #T1120)? A: Yes, both kits employ the same low-volume elution column to enable robust cleanup and concentration of your samples. The Monarch Spin Columns S1A and Tubes are also available for purchase separately (NEB # T2037L). Q: What is the composition of each buffer provided with the Monarch® Spin DNA Gel Extraction Kit (NEB #T1120)? A: The composition of the buffers is proprietary. We can, however, share the following: Monarch Buffer BY: Guanidine thiocyanate dissolving buffer Monarch Buffer WZ: Ethanol-based wash buffer Monarch Buffer EY: 10 mM Tris, 0.1 mM EDTA, pH 8.5 elution buffer Q: What is the maximum binding capacity of the Monarch® Spin Columns S1A provided in the Monarch Spin DNA Gel Extraction Kit (NEB #T1120)? A: The matrix supplied in each column can bind up to 5 μg of DNA. Purifying DNA from an agarose gel can introduce components, such as carbohydrates, that can compete for binding to the matrix and affect DNA recovery. Q: Can I excise a fragment from a gel and store it for purification with the Monarch® Spin DNA Gel Extraction Kit (NEB #T1120) at a later time? A: Yes, the slice can be stored in a closed microfuge tube at 4°C for up to 3 days. However, purification immediately following excision will likely provide the best results. Q: What size DNA can be purified with the Monarch® Spin Columns S1A (NEB #T2037)? A: Recovery of DNA from 50 bp to 25 kb can be achieved with the Monarch Spin Columns S1A, supplied in both the Monarch Spin DNA Gel Extraction Kit (NEB #T1120) and the Monarch Spin PCR & DNA Cleanup Kit (5 µg) (NEB #T1130). Efficiency decreases with longer DNA due to tighter binding to the matrix. Recovery with longer DNA can be increased with modified elution methods such as using heated elution buffer (50°C) and/or incubating at room temperature for 5 minutes after the addition of elution buffer. For detailed instructions, please read the instructions provided in the manual. Q: After purification using the Monarch® Spin DNA Gel Extraction Kit (NEB #T1120), I see a faint additional band running below the expected size on a gel. What could cause that? A: Chaotropic agents used in silica-based DNA purification can induce DNA denaturation and these single-stranded forms of DNA will have faster mobility in a gel. If present, we recommend using salt and heat to renature your sample. Adding NaCl to 10mM and heating the sample to 95°C for one minute followed by a slow cooling to room temperature should promote re-annealing of the denatured strands. Alternatively, if the sample will be used in an enzymatic reaction, the purified DNA can be added to the reaction with the enzyme omitted, heated to 95°C for one minute, and cooled to the reaction temperature followed by adding the enzyme. Q: Are Monarch® Spin Columns S1A (NEB #T2037) compatible with vacuum manifolds? A: We have tested compatibility of Monarch columns from the following Monarch kits for use with common benchtop vacuum manifolds: DNA Gel Extraction Kit (NEB #T1020 & #T1120) PCR & DNA Cleanup Kit (NEB #T1030 & #T1130) RNA Cleanup Kits (NEB #T2030). Please refer to protocols using vacuum manifolds provided in the manual. Vacuum manifolds can be utilized with some other Monarch columns but may require optimization and/or additional time for the liquid to completely flow through. Q: Are the components of the previous version of Monarch® DNA Gel Extraction Kit (NEB #T1020) compatible with the new version? Can I use #T1020 buffers on new Monarch Spin Columns S1A or vice versa? A: Yes, while this new version of the Monarch Spin DNA Gel Extraction Kit (NEB #T1120) is developed with performance improvements, the kit buffers from the previous version are compatible. Using the kit with other Monarch buffers from previous versions will still generate high-quality, purified DNA suitable for various downstream applications. However, there may be up to 20% variance in yield in some cases when not using the supplied, matching buffers within a kit. Q: Are the new, upgraded Monarch® Spin Columns (NEB #T2037) compatible with the QIAcube® system? A: The new Monarch spin columns and collection tubes are engineered to ensure high-quality nucleic acid purification. They feature a low elution volume, no buffer retention, and reduced plastic usage. However, these new columns are not compatible with the QIAcube system. Q: Can I use the Monarch Spin RNA Cleanup Kits to cleanup my in vitro transcription (IVT) reaction? A: Yes, the Monarch Spin RNA Cleanup Kit (50 µg) (NEB #T2040) is suitable for cleaning up in vitro transcription reactions where the yield is not expected to exceed 50 µg. If the RNA yield of an in vitro transcription is expected to exceed 50 µg, we recommend the reaction be split among multiple Monarch Spin RNA Cleanup Columns (50 µg) or cleaned up using the Monarch Spin RNA Cleanup Kit (500 µg) (NEB #T2050), due to the larger RNA binding capacity. Q: I see some silica residue in my Monarch column. Is this a problem? A: No, this is not an issue. The presence of some silica residue or particles in the nucleic acid purification column does not impact nucleic acid binding or purity. Our columns are designed to retain silica fibers, helping ensure they do not end up in the final eluate. Q: Can I use the Monarch Spin DNA Gel Extraction Kit (NEB #T1120) for low DNA input amounts? A: Yes. The Monarch Spin DNA Gel Extraction Kit (NEB #T1120) can be used with DNA inputs as low as 10 ng. An optimized protocol for low-input gel extraction is available in the protocol section. This method is effective for DNA amounts ranging from 10 ng to 500 ng.