{"product_id":"neb-t1135v","title":"New England Biolabs, T1135V, Monarch® Spin High-Capacity DNA Cleanup Kit (100 μg)","description":"\u003cb\u003eRelated Categories\u003c\/b\u003e PCR \u0026amp; DNA Cleanup,, Nucleic Acid Purification  \u003cb\u003eApplications\u003c\/b\u003e Nucleic Acid Purification  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eMaterials Required but not Supplied\u003c\/b\u003e Isopropanol Ethanol  \u003cb\u003eFAQ\u003c\/b\u003e Q: Can I purchase additional Monarch® columns and collection tubes for the Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) (NEB #T1135) separately? A: Yes, columns (NEB #T1157) and collection tubes (NEB #T2118) for Monarch Spin High-Capacity DNA Cleanup Kit 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 S3A (NEB #T1157)? A: The range for elution volume using Monarch Spin Columns S3A with Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) is 50 - 200 μl, with 100 μl as a recommended elution volume. When using minimum elution volume, it is recommended to incubate at room temperature for a few more minutes than recommended to maximize yield. Q: What is the composition of each buffer provided with the Monarch® Spin High-Capacity DNA Cleanup Kit (100 μg) (NEB #T1135)? A: The composition of the buffers is proprietary. We can, however, share the following: Monarch Buffer BZ: Guanidine and isopropanol-based binding 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 S3A provided with the Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) (NEB #T1135)? A: The matrix supplied in each column can bind up to 100 μg of DNA. Purifying DNA from a PCR or other enzymatic reaction introduces components such as detergents, dyes, proteins, and nucleotides, all of which can compete for binding to the membrane and affect DNA recovery. Q: What factors affect the (A260\/230) ratio when using the Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135)? A: Guanidine and ethanol, both introduced during the prep, can reduce the A260\/A230 ratio. Following the protocol will ensure these components are removed. Additionally, though unlikely, some particulates may elute which can affect the ratio as well. If this is the case, an additional 15-second spin of the eluted DNA before evaluation by spectrometry (Nanodrop®) should help. Withdrawing the DNA sample from the top of the sample can help to avoid transferring any particulates. Q: Do you have any recommendations for the purification of single-stranded DNA (ssDNA) when using the Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135)? A: The Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) can be used to purify ssDNA. ssDNA binds less tightly than dsDNA, and the strength of binding will be affected by length, sequence, and the formation of any secondary structure. For ssDNA \u0026lt; 100 nt, we recommend using the Oligonucleotide Cleanup Protocol provided with the kit, which is optimized to recover shorter DNA. While the Monarch Spin PCR \u0026amp; DNA Cleanup Kit (100 μg) will work well with ssDNA, it is difficult to predict the recovery efficiency of ssDNA. We, therefore, recommend a trial recovery with the DNA of interest before using your entire sample. Q: What size DNA can be purified with the Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135)? A: Recovery of DNA from 50 bp to 25 kb can be accomplished with the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) (NEB #T1135). Efficiency decreases with longer DNA due to tighter binding to the matrix. The recovery of DNA ≥ 15 kb can be improved by modifying the elution method, using heated elution buffer (50°C) and extending incubation times. For optimal recovery of large DNA fragments, please follow the instructions in the manual. For DNA \u0026gt; 25 kb, we provide a modified protocol optimized for large DNA on our webpage. Please refer to the protocol available on webpage for detailed procedural guidance. Q: Are Monarch® Spin Columns S3A (NEB #T1157) compatible with vacuum manifolds? A: Yes, Monarch Spin Columns S3A are compatible with vacuum manifolds. 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: Can I use the Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135) to purify oligonucleotides and other DNA fragments? A: Yes. A simple protocol modification of adding ethanol to the sample before binding increases the binding efficiency of small DNA to the matrix, allowing oligos and short dsDNA to bind to the column. Please refer to the Oligonucleotide Cleanup Protocol. Q: Can I use Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135) to purify circular DNA? A: Yes, you can use the Monarch Spin High-Capacity DNA Cleanup Kit (NEB #T1135) to purify circular DNA, including both single-stranded (ssDNA) and double-stranded (dsDNA), by following the standard DNA cleanup protocol. Q: Can I use Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135) to purify DNA \u0026gt; 25 kb or genomic DNA? A: Yes, you can use the Monarch Spin High-Capacity DNA Cleanup Kit (NEB #T1135) to purify large DNA, including genomic DNA or RCA product (Rolling Circle Amplification), by following the modified protocol provided. Please refer to the corresponding protocol on our webpage for detailed instructions. Q: Are the Monarch® Spin Columns S3A (NEB #T1157) 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: 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. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835509100713,"sku":"T1135V","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-t1135v","provider":"Iright","version":"1.0","type":"link"}