{"product_id":"neb-t3010l","title":"New England Biolabs, T3010L, Monarch® Spin gDNA Extraction Kit","description":"Effective November 20, 2025, protocols have been updated to remove the low-speed spin step (1000 × g), streamlining the workflow and saving time. Our testing has shown no impact on performance or yield. Kits using orange-tinted Monarch Spin Columns S2C (  \u003cb\u003eRelated Categories\u003c\/b\u003e Genomic DNA Extraction \u0026amp; Purification,, Nucleic Acid Purification  \u003cb\u003eApplications\u003c\/b\u003e Nucleic Acid Purification  \u003cb\u003eFAQ\u003c\/b\u003e Q: What is the composition of each buffer provided with the Monarch® Spin gDNA Extraction Kit? A: The compositions of the buffers are proprietary. However, we can share the following information: Monarch® gDNA Tissue Lysis Buffer....................Detergent-based lysis buffer Monarch® gDNA Cell Lysis Buffer........................Guanidinium hydrochloride-based lysis buffer Monarch® gDNA Blood Lysis Buffer.....................Guanidinium hydrochloride-based lysis buffer Monarch® gDNA Binding Buffer...........................Guanidinium thiocyanate-based buffer Monarch® gDNA Wash Buffer..............................Ethanol-based wash buffer Monarch® gDNA Elution Buffer............................10 mM Tris-HCl, pH 9.0, 0.1 mM EDTA Q: What is the maximum binding capacity of the Monarch® Spin gDNA Extraction Kit columns? A: The column matrix will bind 25-30 µg of genomic DNA (gDNA), although in some cases greater yields have been observed. It is important to note that the binding capacity of the matrix will be reduced if significant amounts of insoluble proteins or polysaccharides are present in the lysate.Please refer to our guidelines for choosing input amounts to avoid overloading the columns. Additionally, if the gDNA is longer than 80 kb, it cannot be efficiently eluted from the matrix. Q: What is the minimum elution volume that can be used with the Monarch® Spin gDNA Extraction Kit columns? A: The minimum elution volume is 35 µl. For maximum recovery, 100 µl is recommended. Smaller elution volumes will lead to more concentrated samples but also a lower yield. Typically, when 35 µl is used for elution, yields are approximately 20% lower than when using 100 µl. Using elution volumes lower than 35 µl will result in inconsistent wetting of the silica matrix and significantly reduced recovery. Recovery of Genomic DNA using various elution volumes in the Monarch Spin gDNA Extraction Kit Optimal elution volume range for elution is 35 – 100 µl. A lysate pool was prepared by using 10 mg RNAlater®-stabilized rat liver samples and following the Monarch tissue protocol. Elution volumes used in triplicates samples were 25, 30, 35, 40, 50, 75, and 100 µl. The average yield obtained with 100 µl elution volume was 19.1 µg, which was considered 100% yield. Using 35 µl to elute, the average yield was 81.6%; 25 µl and 20 µl yielded 74% and 70.6%, respectively. For volumes ≥35 µl, the recovered volume after elution was ~5 µl lower than the elution volume added and for \u0026lt;30µl the recovered volume was ~10 µl lower. Tests performed with other starting materials showed similar results, with a 20-25% reduction in overall yield when elution volumes were reduced from 100 to 35 µl. For more information, see “Considerations for Elution and Storage” in the product manual. Q: How important is it to pre-heat the elution buffer to 60°C? Would 56°C be OK? A: The pre-heating of the elution buffer is essential for maximal recovery with the Monarch® Spin gDNA Extraction Kit. If more convenient, a temperature of 56°C can be used but may result in a very slight reduction in yield. Room temperature elution buffer can also be used but will result in a 20-30% reduction in yield and may require a second elution. Additionally, at room temperature, large gDNA fragments will elute less efficiently, so the eluted gDNA will have a smaller average size. For more information, see “Considerations for Elution and Storage” in the product manual. Q: Can I get better recovery with the Monarch® Spin gDNA Extraction Kit if I do a second elution with my eluent from the first elution? A: The Monarch protocol for gDNA purification has been optimized for maximum speed and convenience. The elution step with preheated elution buffer enables maximal recovery in just one elution step. However, a second elution step can be performed to maximize recovery. This second elution can be performed using a fresh aliquot of Monarch gDNA Elution Buffer or the eluent from the first elution, to minimize dilution of the DNA. A 10% increase in the total yield may be achieved. For more information, see “Considerations for Elution and Storage” in the product manual. Q: Can I use a different elution buffer than the one supplied with the Monarch® Spin gDNA Extraction Kit (NEB #T3010)? A: The elution buffer supplied with the kit is 10 mM Tris-HCl, pH 9.0, 0.1 mM EDTA. This buffer is optimal for long term storage of gDNA. Any low-salt buffer can be used for elution, such as 10 mM Tris-HCl pH 8.5, TE Buffer or nuclease-free water. Preheating the elution buffer to 60°C ensures rapid and efficient elution of the gDNA. For more information, see “Considerations for Elution and Storage” in the product manual. Q: What size gDNA can be purified with the Monarch® Spin gDNA Extraction Kit? A: Assuming high quality input material has been used, the peak size of gDNA isolated with this kit ranges from 50-60 kb. Typical DNA Integrity Numbers (DINs) for gDNA isolated from blood, cells or tissues are around 9.0. The Monarch gDNA purification protocol is optimized for isolation of gDNA that is larger than that isolated by other commercial kits. For example, it includes elution with preheated buffer that enhances the elution of larger pieces of DNA. Some starting materials such as buccal swabs or saliva, however, contain a significant proportion of dead cells. In those cells, apoptotic mechanisms have activated nucleases that digest the gDNA to shorter fragments. The peak size of gDNA isolated from such samples is usually in the range 20-40 kb. Q: What is the minimal input amount that can be used with the Monarch® Spin gDNA Extraction Kit? A: Input material amounts that result in expected yields of around 100 ng of gDNA correspond to: 5 µl whole blood, 1 x 104 cultured cells or 0.2 mg tissue. If a smaller amount of input material is used, non-specific binding to surfaces will cause the recovery rate to drop. For best results with very small amounts of input material, the use of carrier RNA is recommended. See the product manual for more details. Q: Can I use the Monarch® Spin gDNA Extraction Kit to clean up gDNA that was extracted with phenol\/chloroform? A: Yes, the Monarch Spin gDNA Extraction Kit can be used to clean up phenol\/chloroform purified gDNA by following the protocol for Genomic DNA Cleanup. However, recovery may be lower than the usual 80% because phenol\/chloroform purified DNA typically yields longer fragments. Although the use of preheated elution buffer in the Monarch protocol facilitates the elution of large gDNA fragments, the fraction of gDNA that is longer than 80 kb will be eluted less efficiently from the silica matrix. Q: Can I use the Monarch® Spin gDNA Extraction Kit for gDNA cleanup? A: Yes, the Monarch Genomic DNA Purification Kit can be used for cleaning up gDNA from enzymatic reactions or previous purification protocols by following the protocol for Genomic DNA Cleanup. Cleanup can be performed either directly on the gDNA or with a prior enzyme inactivation step using Proteinase K. Recovery of approximately 80% gDNA may be expected with 1 elution step when the input amount is \u0026gt;100 ng. However, if the input DNA is longer than 80 kb, which may be the case with gDNA that was purified following, for example, phenol-chloroform or salting-out protocols, the elution efficiency may be lower because gDNA \u0026gt;80 kb is not eluted efficiently from the silica matrix. Q: How can I assess gDNA integrity and purity? A: Purity of eluted gDNA samples is assessed by reviewing OD ratios collected during routine spectrophotometry. Pure gDNA typically has an A260\/280 of 1.8–1.9 and an A260\/230 of 1.8–2.5. The integrity of the gDNA is assessed by loading approximately 100 ng per sample on a 0.75% agarose gel and comparing size distribution to a suitable marker, such as lambda DNA, either as full length DNA (NEB #N3011) or digested with HindIII (NEB #N3012). Typically, for intact gDNA, the majority of the gDNA signal will be larger than the upper band of the HindIII digest. Alternatively, gDNA samples can be run on an Agilent TapeStation® using a Genomic DNA ScreenTape. This system will provide information on the peak size of the gDNA and the overall integrity via the DIN (DNA Integrity Number). High peak sizes (\u0026gt;50 kb) and DINs \u0026gt;8.5 indicate that the DNA is of high quality. For more information on assessing nucleic acid purity, download our application note, A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers. Q: Can I use a different RNase A with the Monarch® Spin gDNA Extraction Kit? A: As long as the RNase A has a similar concentration and activity specifications, it should be OK to use. However, many RNase A products do not have the same quality specifications as Monarch RNase A, which has no detectable endonuclease, exonuclease, DNase and protease activity and is free of DNA. These specifications may be relevant for your downstream applications. Q: Can I use a different Proteinase K with the Monarch® Genomic DNA Purification Kit? A: As long as the Proteinase K has a similar concentration and activity specifications, it should be OK to use. 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. However, many other Proteinase K products do not have the same specifications as NEB’s Proteinase K products (NEB #P8200 or #P8107), which has been highly characterized for more consistent performance. Additionally, NEB Proteinase K has no detectable endonuclease, exonuclease and DNase activity and is DNA-free. These specifications may be relevant for your downstream applications. Q: Can I omit the low-speed binding centrifugation step to save time? A: You may omit this step if you wish and you will still get good results. However, results will be inconsistent, and depending on the starting material, yields may be reduced by 5-30%. Binding at low speed is particularly important for optimal recovery when processing tissue samples containing high amounts of DNA and when carrying out the Genomic DNA Cleanup Protocol. Q: Can I omit the RNase A digestion step from the protocol? A: Yes, the Monarch gDNA purification procedure has been designed to minimize the RNA amount that is co-purified. Typical percentages of RNA that are found in gDNA Monarch eluates when the RNase A digestion step is omitted are 1% (Blood Protocol), 10% (Cell Protocol), and 1-5% (Tissue Protocol). These low amounts typically do not affect most downstream applications. If the RNase A digestion step is included in each protocol, final RNA amounts that are co-purified with the gDNA are 0-1%, amounts that are not detectable by most methods. Q: How should I store purified gDNA samples? A: Purified gDNA samples can be stored at 4°C. However, for long term storage, we recommend storing them at -20°C. The Monarch® Spin gDNA Extraction Kit (10 mM Tris-HCl, pH 9.0, 0.1 mM EDTA) that is supplied with the Monarch Genomic DNA Purification Kit provides extra safety for long term storage of gDNA samples. Q: Is Monarch-purified gDNA suitable for long-range PCR? A: Yes. The large fragment size of Monarch-purified gDNA makes it excellent starting material for long range PCR approaches. For long range PCR, we recommend LongAmp TaqDNA Polymerase (NEB #M0323), LongAmp Taq 2x Master Mix (NEB #M0287) or LongAmp Hot Start Taq Polymerase (NEB #M0534). Genomic DNA purified with the Monarch Spin gDNA Extraction Kitis high quality and suitable for sensitive applications like long range PCR and qPCR A. Amplification reactions were set up with primer pairs specific for 6, 8, 10, 12, 16, 20 kb amplicons from human DNA. LongAmp® Hot Start Taq 2x Master Mix (NEB #M0533) was used and 25 ng template DNA was added to each sample. PCR reactions were carried out on an Applied Biosystems 2720 Thermal Cycler. Monarch-purified genomic DNA isolated from HeLa cells and human blood were compared to commercially available reference DNA from the human cell line NA19240 F11. 10 of 20 μl was loaded on a 1.5% agarose gel, using the 1 kb DNA Ladder (NEB #N3232) as a marker. Results indicated DNA was of high-integrity and suitable for long range PCR. B. Monarch-purified genomic DNA from human whole blood, HeLa cells and mouse tail was diluted to produce a five log range of input template concentrations. The results were generated using primers targeting gHEME (human whole blood) and gREL (HeLa, mouse tail) for qPCR assays with the Luna Universal qPCR Master Mix (NEB #M3003) and cycled on a Bio-Rad® CFX Touch qPCR thermal cycler. Results indicated that DNA is highly pure and free from inhibitors, optimal for qPCR. Q: Can I purchase Monarch® buffers and columns separately? A: Monarch spin columns and several Monarch buffers and reagents are also offered separately. Q: When working with other commercial silica column-based kits, I occasionally see a white precipitate in the eluate. What is this? A: Some silica matrices release silica fibers during the elution step. Although silica fibers generally do not affect most downstream applications, they influence OD measurements and should be removed by centrifugation if reliable spectrophotometric values are needed. Monarch Genomic DNA Purification Columns contain a retaining membrane that prevents silica fibers from passing through, thus eliminating the need to perform additional centrifugation prior to accurate spectrophotometric analysis. Q: Can I use Monarch® RNase A for applications\/workflows other than gDNA purification? A: Although NEB has not specifically validated the use of Monarch RNase A in applications outside of gDNA purification, the enzyme should work well for other applications that utilize RNase A. Q: Is Monarch-purified gDNA compatible with Next Generation Sequencing (NGS) platforms like PacBio and nanopore? A: Yes. gDNA purified with this kit can be used for various applications on these sequencing platforms. Assuming high quality input material has been used, read lengths of up to 50 kb can be routinely achieved using Monarch purified gDNA from different starting materials. Monarch purified DNA quality is superior to SDS Proteinase K-purified DNA commonly used for Nanopore sequencing 2 µg E. coli genomic DNA extracted using either Monarch or by following a traditional SDS Proteinase K isopropanol protocol was enzymatically fragmented with identical incubation times and used as input DNA for MinION library preparation. Oxford Nanopore Technology’s 1D kit protocol was followed to prepare libraries. Both libraries were run on the MinION, the data collected for 48 hours and base called using Albacore. Monarch-purified DNA performed better on all sequencing metrics, including read length, read quality and total sequencing data collected. The graphs illustrate that Monarch purified data provides a higher percentage of high quality sequencing data than SDS Proteinase K purified DNA. Monarch provides excellent quality starting material for Pacific Biosciences (PacBio) sequencing Bacterial gDNA was isolated from 5x109 E. coli ER2683 cells with pACYC184 plasmid by following the Monarch rapid protocol for Gram- bacteria. 10 µg Monarch-purified gDNA was sheared with a Covaris® g-TUBE, targeting 10 kb fragment size. 5 µg sheared DNA was used for library preparation according to the standard PacBio protocol (without size selection). Samples were sequenced on a Pacific Biosciences RSII sequencer, using the P6\/C4 chemistry, loaded as 10 pM SMRTbell library and data were collected in a 5 hr movie. Total bases (yield): 1208 Mb, Average Polymerase read length: 16,046, Polymerase read quality: 0.85. Average Reads of Insert length: 6,060, Reads of Insert quality: 0.89, Longest read of insert: 43,680. A. Insert lengths indicate that the majority of the reads were in the desired insert size range. B. The polymerase read lengths indicate that the purified DNA was of high quality to allow the enzyme to read the insert several times within the SMRTbell construct. C. The loading evaluation data further illustrates that the purified DNA is of high quality. Q: Why did the silica membrane on my Monarch Spin Columns S2C (NEB #T3017) dislodge during my genomic DNA prep? A: Monarch Spin Columns S2C were designed without a retaining ring in order to prevent contamination from buffer carryover and ensure maximum purity of eluted DNA. When the protocol is carried out as specified, the membranes are held in place and should not dislodge. However, the membrane can expand and dislodge under very specific conditions: The column is loaded more than once The lysed sample or the wash buffer is allowed to sit on the membrane for too long The column is vortexed The input amounts are exceeded significantly The membrane could be more likely to dislodge during low-speed spins. For kits containing orange-tinted Monarch Spin Columns S2C (NEB #T3017), follow the current protocols, which do not use a low-speed spin (1,000 x g for 3 minutes) Accordingly, we recommend that the protocol is followed as specified and that the column is never reloaded or vortexed. Q: Can the Monarch® Spin gDNA Extraction Kit (NEB #T3010) be used to isolate gDNA from drosophila and other insects? A: Yes, and we recommend following the detailed protocol for Genomic DNA Purification from Insects. This is a modified version of the protocol for genomic DNA purification from tissues, optimized for processing insect samples. Q: Can the Monarch® Spin gDNA Extraction Kit (NEB #T3010) be used to isolate gDNA from plants? A: Although not specifically optimized for this application, many users have had success processing plant samples (cannabis, wheat leaf, orchid leaf) with the Monarch Spin gDNA Extraction Kit. However, plant samples contain complex polysaccharides and polyphenols, neither of which the Monarch buffer system has been optimized to address. We recommend following the tissue protocol. Before lysis with the Tissue Lysis Buffer, the plant cell walls should be opened as much as possible by either using a microtube pestle, grinding under liquid nitrogen or using a bead mill. After lysis and RNase A incubation, centrifuge for 3 minutes at maximum speed (≥12,000 x g) to remove solid material and transfer the supernatant to a fresh tube. We recommend using \u0026lt;25 mg of starting material, and using young tissue is also recommended for best results. Yields will vary significantly amongst different plant species based on both their genome size and the accessibility of the DNA. Q: Can the Monarch® Spin gDNA Extraction Kit (NEB #T3010) be used to isolate gDNA from plasma, serum and urine? A: We do not recommend this kit for isolation of genomic DNA from plasma, serum or urine. This kit has not been validated on these sample types, and is not optimized for processing large volumes of starting material. Q: Can the Monarch® Spin gDNA Extraction Kit (NEB #T3010) be used for isolating cfDNA? A: We do not recommend this kit for isolation of cfDNA. cfDNA contains small DNA fragments, and the binding chemistry in this kit is not optimized for small fragments. Additionally, the workflow is not optimized to process large volumes of starting materials, which is often necessary when isolating cfDNA. Q: Can the Monarch® Spin gDNA Extraction Kit (NEB #T3010) be used with a vacuum manifold? A: Yes, the kit can be used for with a vacuum manifold for the binding and first wash step of Part 2: Genomic DNA Binding and Elution. The second wash step and the elution should be carried out in a centrifuge. Purity of vacuum-purified samples may be reduced, as protein and other cell components can remain on the membrane; high speed centrifugation is recommended for maximum purity and reproducibility. Q: The enzymes in the kit were not stored at -20°C right away. Will they still work? A: Yes, unopened Monarch Proteinase K and RNAse A enzymes are stable at ambient temperatures short-term. After opening, Proteinase K and Monarch RNAse A should be stored at -20°C. Q: Does it matter which anticoagulant is used in blood samples? A: No. gDNA isolation from blood with the Monarch® Spin gDNA Extraction Kit works well for blood that is treated with either EDTA, citrate or heparin. Q: There is a precipitate in the Proteinase K enzyme tube, is this normal? A: It is not uncommon to see a white, amorphous precipitate in the Proteinase K tube. There is no impact to Proteinase K activity or kit function, so you can proceed as normal with your workflow. 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: I noticed that in the Monarch® Spin gDNA Extraction Kit (NEB #T3010), the Monarch Spin Columns S2C (NEB #T3017) for gDNA purification are now tinted orange. Does this impact performance? A: The tinted color does not impact performance. Follow the recommended usage guidelines, including maximum input guidelines in the manual. Store the tinted columns protected from light to avoid fading of the tint over time. There may be natural variations in the tinting across the columns. Q: Why was the low-speed spin (1,000 x g for 3 minutes) removed from the protocols in the Monarch Spin gDNA Extraction Kit (NEB #T3010)? Can I still perform this low-speed spin? A: The protocol change to remove the low-speed spin step (1,000 x g for 3 minutes) in the binding step is for streamlining the workflow and saving time. Our testing has shown no impact on performance or yield. Kits using orange-tinted Monarch Spin Columns S2C (NEB #T3017) are encouraged to follow the updated protocols for improved efficiency. 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. 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":46835481018537,"sku":"T3010L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-t3010l","provider":"Iright","version":"1.0","type":"link"}