New England Biolabs, T1130S, Monarch® Spin PCR & DNA Cleanup Kit (5 μg)

Related Categories PCR & DNA Cleanup,, 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: What is the composition of each buffer provided with the Monarch® PCR & DNA Cleanup Kit (5 μg) (NEB #T1130)? 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 S1A provided with the Monarch Spin PCR & DNA Cleanup Kit (5 μg) (NEB #T1130)? A: The matrix supplied in each column can bind up to 5 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 PCR & DNA Cleanup Kit (NEB #T1130)? 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: What size primers can be effectively removed from a PCR reaction using the Monarch® Spin PCR & DNA Cleanup Kit (NEB #T1130)? A: Primers up to 45 nucleotides in length are most efficiently removed using the standard conditions recommended in the kit. Longer primers may also be removed, but any secondary structure or complementarity to other DNA species in the mix may increase initial binding to the matrix and result in trace recovery. Q: Can the Monarch® Spin PCR & DNA Cleanup Kit (5 μg) (NEB #T1130) be used to purify RNA? A: RNA will bind to the Monarch Spin Column S1A, but the buffers and protocol have not been optimized to ensure reliable, RNase-free recovery. Therefore, we do not recommend using this kit to purify RNA. NEB offers 3 kits designed specifically for the cleanup and concentration of RNA samples (NEB #T2030, #T2040, #T2050). Q: Do you have any recommendations for the purification of single-stranded DNA (ssDNA) when using the Monarch® Spin PCR & DNA Cleanup Kit (NEB #T1130)? A: The Monarch Spin PCR & DNA Cleanup Kit (5 μg) can be used to purify ssDNA from enzymatic reactions, as well as for general cleanup to remove potential inhibitors of PCR. 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 < 200 nt, we recommend using the Oligonucleotide Cleanup Protocol provided with the kit, which is optimized to recover shorter DNA. While the Monarch Spin PCR & DNA Cleanup Kit (5 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: 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 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 accomplished 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. 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. 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: Can I use the Monarch® Spin PCR & DNA Cleanup Kit (NEB #T1130) 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: Are the components of the previous version of Monarch® PCR & DNA Cleanup Kit (NEB #T1030) compatible with the new version? Can I use #T1030 buffers on new Monarch Spin Columns S1A (NEB #T2037) or vice versa? A: Yes, while the new version of the Monarch Spin PCR & DNA Cleanup Kit (NEB #T1130) 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% variances 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: 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.