{"product_id":"neb-t2047l","title":"New England Biolabs, T2047L, Monarch® Spin Columns S2A","description":"  \u003cb\u003eRelated Categories\u003c\/b\u003e RNA Cleanup,, RNA Extraction and Purification,, Nucleic Acid Purification  \u003cb\u003eFAQ\u003c\/b\u003e Q: I notice that I'm applying force to fit the column into a collection tube. Should I be doing this? What collection tubes are compatible with columns? A: Matching collection tubes are provided in all Monarch® kits for the spin columns. If you use collection tubes outside of a given kit, please use the Monarch® Spin Collection Tubes (NEB #T2118). Do not force any columns into non-recommended collection tubes that have a very tight fit to avoid damaging the column. Q: Can the Monarch Spin RNA Cleanup Kits be used for RNA extraction? A: Yes. Although originally developed for RNA cleanup applications, these kits can also be used for RNA extraction from cells, saliva, and swab samples. When used for viral extraction, the Monarch DNA\/RNA Protection Reagent (sold separately, NEB #T2011) must be used for viral inactivation. Visit our online protocols for more information: RNA Extraction from Cells Using the Monarch Spin RNA Cleanup Kits RNA Extraction from Buccal\/Nasopharyngeal Swabs Using the Monarch Spin RNA Cleanup Kits RNA Extraction from Saliva Using the Monarch Spin RNA Cleanup Kits Automation of Viral RNA Extraction from Saliva Using the Monarch Spin RNA Cleanup Kit Q: What factors affect my (A260\/A230) when using the Monarch Spin RNA Cleanup Kits? A: Guanidine and ethanol, both introduced during the prep, can reduce the A260\/A230 ratio. Following the protocol, especially the wash steps, will ensure these components are removed. Q: Can I purchase Monarch® buffers and columns separately? A: Monarch spin columns and several Monarch buffers and reagents are also offered separately. 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: Are the columns in the Monarch Spin RNA Cleanup Kits the same as those in the Monarch Total RNA Miniprep Kit (NEB #T2010)? A: No, the columns in the Monarch RNA Cleanup Kits (NEB #T2030, #T2040 and #T2050) are not the same as the ones in the Monarch Total RNA Miniprep Kit (NEB #T2010). Q: Can I use the Monarch Spin RNA Cleanup Kit to cleanup up my DNase I-treated RNA? A: Yes, the Monarch Spin RNA Cleanup Kits will remove residual DNase I (NEB #M0303) activity from RNA samples. On-column treatment with DNase I can often result in residual DNase I activity, therefore, we do not recommend on-column DNase I treatment with the Monarch Spin RNA Cleanup Kit, but rather in-solution DNase I treatment prior to RNA cleanup. Q: Are the Monarch Spin RNA Cleanup Kits compatible with NEBNext reagents for RNA library prep? A: Yes. RNA purified with this kit can be used for RNA library preparation using NEB's NEBNext RNA Library Prep reagents. Q: What is the maximum binding capacity of the column for RNA Cleanup Column provided with the Monarch Spin RNA Cleanup Kit? A: Although in some cases greater yields have been observed, the matrix supplied in each column is sufficient to bind the following quantities of RNA: T2030\/T2037: 10 µg of RNA T2040\/T2047: 50 µg of RNA T2050\/T2057: 500 µg of RNA It is important to note that enzymatic reactions introduce components such as detergents, dyes, protein and nucleotides, all of which can compete for binding to the membrane during cleanup and affect RNA recovery. Q: Can I get better recovery with the Monarch Spin RNA Cleanup Kits if I do a second elution with my eluent from the first elution? A: A second elution can be performed to maximize RNA recovery. This second elution can be performed using a fresh aliquot of nuclease-free water or the eluent from the first elution to maximize recovery while minimizing volume. Q: Can I use the Monarch Spin RNA Cleanup Kits to purify RNA from agarose gels? A: Yes, the Monarch Spin RNA Cleanup Kit manual contains a protocol which will purify RNA from agarose gels, though this is not their primary application. Recoveries from extraction of RNA from agarose range from 40-70%, which is less than expected recovery for RNA Cleanup from enzymatic reactions. The protocol can also be accessed online. Q: What size RNA can be purified with the Monarch Spin RNA Cleanup Kit? A: Using the standard protocol, RNAs ≥ 25 nt can be purified. With a slight modification in the protocol (adding 2 volumes of ethanol instead of 1 volume after addition of the binding buffer), RNA \u0026gt; 15 nt can be purified. Recovery of Small RNAs using the Monarch Spin RNA Cleanup Kit. Synthesized RNA oligonucleotides (1 μg in 50 μl H2O) of varying lengths (15–100 nt) were purified using the Monarch Spin RNA Cleanup Kit (NEB #T2040) and eluted in 50 μl nuclease-free water. The percent recovery of Spin RNA was calculated from the resulting A260, measured using a Trinean DropSense™ 16. The Monarch Spin RNA Cleanup Kit standard protocol results in \u0026gt;70% recovery and cleanup of RNA ≥ 25 nt, while use of the Monarch Spin RNA Cleanup Kit with the modified protocol (addition of 2 volumes of ethanol in step 2) results in \u0026gt;70% recovery and cleanup of even smaller RNAs (as low as 15 nt). Please note that recovery of small RNAs (\u0026lt; 45 nt) can be affected by sequence, interactions with other nucleic acids and\/or secondary structure. Secondary structure affects recovery of small RNAs (\u0026lt;45 nt) Synthesized RNA oligonucleotides (1 μg in 50 μl H2O) of varying lengths (25-40 nt) and varying predicted secondary structure* were purified using the Monarch Spin RNA Cleanup Kit (NEB #T2040) and eluted in 50 μl nuclease-free water. The percent recovery of the RNA was calculated from the resulting A260 as measured using a Trinean DropSense™ 16. Small RNAs (25–40 nt) with low predicted secondary structure were recovered efficiently using the Spin Monarch RNA Cleanup Kit standard protocol. However, increased levels of secondary structure decrease the recovery of small RNAs (25–40 nt in length). Recovery can typically be increased to \u0026gt;70% (orange line) using the Spin Monarch RNA Cleanup Kit with the modified protocol (addition of 2 volumes of ethanol in step 2). *Secondary structure was predicted using RNAstructure Web Server: Reuter, J. S., \u0026amp; Mathews, D. H. (2010). RNAstructure: software for RNA secondary structure prediction and analysis. BMC Bioinformatics. 11,129 If poor yield of a small RNA is observed using the standard RNA Cleanup Protocol (adding 1 volume of ethanol prior to binding to the column), we recommend using the modified protocol (adding 2 volumes of ethanol prior to binding to the column), which uses a higher ethanol-to-sample ratio to shift the exclusion cut-off and enable the capture of smaller RNAs. Q: I would like to clean up  ~150 µg of RNA. Would it be better to use multiple 50 µg columns (NEB #T2040\/#T2047) or a single 500 µg column (NEB #T2050\/#T2057)? A: We recommend using a 500 µg column (NEB #T2050\/#T2057) for the purification of 150 µg of RNA rather than 3-4 of the 50 µg columns. We would expect at least 80-90% recovery of 150 µg on the 500 µg column and it would be more cost effective than using several of the lower capacity columns. It is worth noting, however, that the 500 µg column requires a 5 minute incubation with nuclease-free water during elution, so the workflow is slightly longer. Q: Are the Monarch Spin RNA Cleanup Kits compatible with Luna RT-qPCR reagents? A: Yes. RNA purified with these kits can be used for RT-qPCR with all of NEB’s Luna One-Step RT-qPCR Kits and the LunaScript® RT products. Q: Can the Monarch Spin RNA Cleanup Kits (NEB #T2030, #T2040, #T2050) also be used to purify DNA? A: We have successfully used the Monarch Spin RNA Cleanup Kits for the purification of RNA, ssDNA and dsDNA. However, for optimal cleanup of DNA, we recommend the use of our Monarch Spin PCR \u0026amp; DNA Cleanup Kit (NEB #T1130). 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: Can I use the Monarch Spin RNA Cleanup Kits to cleanup RNA after a TRIzol®\/chloroform extraction? A: Yes, the Monarch Spin RNA Cleanup Kit can be used to clean up the aqueous phase from any guanidinium thiocyanate-phenol-chloroform extraction eliminating the need for tedious RNA precipitation steps. The protocol can be accessed online or in the product manual. Q: Can I do an on column DNase I treatment with the columns for RNA Cleanup? A: DNase I will not be completely removed if used for on-column digestion with the Monarch columns for RNA Cleanup (Monarch Spin Columns S1A, S2A, S2B). In-solution DNase I treatment is recommended, followed by RNA cleanup. Q: What is the smallest volume of nuclease-free water that can be used for elution with the Monarch Spin Columns S1A, S2A, S2B for RNA Cleanup? A: T2030\/T2037: 6 µl; typical elution volumes are 6-20 µl T2040\/T2047: 20 µl; typical elution volumes are 20-50 µl T2050\/T2057: 50 µl; typical elution volumes are 50-100 µl Using lower elution volumes will likely result in variance due to inconsistent wetting of the matrix. Yield may slightly increase if a larger volume is used, but the RNA will be less concentrated. *A second elution can be performed to maximize RNA recovery. This second elution can be performed using a fresh aliquot of nuclease-free water or the eluent from the first elution to maximize recovery while minimizing volume. Recovery of RNA from Monarch Spin RNA Cleanup Kits with Varying Elution Volumes rRNA (10, 50 or 500 µg, respectively of 16S and 23S Ribosomal Standard from E. coli, Sigma) was purified using a Monarch Spin RNA Cleanup Kit (10 µg, NEB #T2030) (50 µg, NEB #T2040) (500 µg, NEB #T2050). Nuclease-free water was used to elute the RNA. The percent recovery of the RNA was calculated from the resulting A260 as measured using a Trinean DropSense 16. ~80% of RNA can be efficiently recovered in 6 µl from the Monarch Spin RNA Cleanup Kit (10 µg, NEB #T2030), 20 µl from the Monarch Spin RNA Cleanup Kit (50 µg, NEB #T2040), and 50 µl from the Monarch Spin RNA Cleanup Kit (500 µg, NEB #T2050). Q: Is the Monarch Spin RNA Cleanup Kit (NEB #T2040) compatible with the EnGen sgRNA Synthesis Kit, S. pyogenes? A: Yes, sgRNAs synthesized using the EnGen sgRNA Synthesis Kit, S. pyogenes (NEB #E3322) can be cleaned up using the Monarch Spin RNA Cleanup Kit (50 µg, NEB #T2040) prior to the formation of ribonucleoprotein (RNP) complexes and subsequent electroporation into cells. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835537641641,"sku":"T2047L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-t2047l","provider":"Iright","version":"1.0","type":"link"}