High Quality Plasmid Purification
The Monarch Spin Plasmid Miniprep Kit offers a unique method for plasmid preparation and purification, enabling extraction of highly concentrated plasmid DNA from various bacterial strains, with yields up to 20 μg of high-quality DNA. The Kit features upgraded spin columns precision engineered to uniquely allow for low elution, in as little as 30 μl, for highly concentrated DNA in downstream application such as sequencing, restriction digestion, transformation, transfection, and more.
A colored buffer system helps indicate completion of key lysis and neutralization steps for an easy to follow workflow and shorter, faster protocols allow for reduced hands-on time and less spin time.
Protokoll-Video Monarch Plasmid Miniprep
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Designed with sustainability in mind
The kit offers significantly less plastic through uniquely designed columns and bottles with plastic reduction while still maintaining structural integrity for centrifugation or vacuum manifolds and other steps.
Kit boxes, inserts, and protocol cards are made of recycled paper.
Unique Column Design
NEB Monarch’s unique column design and membrane assembly allows high-quality, highly-concentrated nucleic acid purification with low elution volume, for downstream applications. The column is designed and made with significantly less plastic for a reduced environmental impact.
Isolate high-quality, concentrated plasmid DNA
Optimized Buffer System
Colored buffers prevent confusion and allow for monitoring of certain steps in the protocol. RNase is provided as part of the neutralization buffer, so there is no need to add it before starting, and no risk of forgetting to add it.
Monarch Spin Plasmid Miniprep Kit consistently produces more concentrated plasmid DNA with equivalent or better yield and purity compared to the leading supplier.
Preps were performed according to recommended protocols using ~1 ml (OD600 = 3) aliquot for pUC19 and pHT01 and ~2 ml (OD600 = 6) aliquot for Rluc and pXba of the same overnight cultures.
Dos and Dont’s of Plasmid Minipreps
Are you planning to perform some plasmid minipreps? Don’t stress! Here are some basic things to keep in mind in order to get clean plasmid DNA, ready for use in downstream applications.
Video: Tipps für die Plasmid Miniprep
DON’T use too many cells
If the recommended amount of cells is exceeded, the amount of lysis buffer recommended in our Monarch Plasmid Miniprep Kit protocol may not be able to efficiently lyse all the cells. Also, excess cell debris resulting from lysis of too many cells can clog the column. If you need to use more cells than recommended, consider splitting the sample in half and using two columns.
DO lyse your cells completely
In order to release ALL of the plasmid DNA, ALL of the cells need to be lysed. To do this, make sure the cells are resuspended completely, without any clumps, and incubate the cells for the recommended amount of time.
DON’T vortex your cells after lysis
Vortexing can cause shearing of host chromosomal DNA, resulting in gDNA contamination.
DON’T skip or shorten the RNase A digestion step
Lucky for you, Monarch Neutralization Buffer comes with RNase A already added (other kits require you to add it – an extra step that is easy to forget!). The neutralization step is very important, as this is the time when RNase A digests the contaminating RNA. It is important to follow the incubation recommendations for this step to ensure complete RNA removal. If >2.5 ml of cell culture is used, increasing the spin time after neutralization to 5 minutes will help.
DO use both wash buffers as directed
Both Monarch wash buffers should be used in the volumes recommended to ensure removal of cell debris, endotoxin and salts.
DON’T mix up your buffers
The buffers need to be added in a particular order, since each one carries out a different function in the purification workflow. Using them out of order can cause your miniprep to fail. Monarch miniprep buffers are color coded for your convenience.
DON’T let the tip of the column touch the flow-through in the collection tube after washing
Ethanol can carry over from the collection tube to the column tip. Ethanol in your eluate can interfere with downstream applications. If you suspect that the tip has touched the flow-through, another spin should do the trick.
DO heat the elution buffer when purifying large plasmids >10kb
Large DNA binds more tightly to the silica matrix. Heating the elution buffer before applying to the column helps to more efficiently release the DNA from the matrix.
Monarch Spin Plasmid Miniprep Products:
| PRODUCT | Prod.Nr.: | SIZE | INFO | |
|---|---|---|---|---|
| Monarch Spin Plasmid Miniprep Kit | T1110S | 50 preps |  |
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| Monarch Spin Plasmid Miniprep Kit | T1110L | 250 preps | |
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| Monarch Spin Columns S2D and Tubes | T1117L | 100 preps | |
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| Monarch Spin Collection Tubes | T2118L | 100 preps | |
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Stand: 05.07.2024
Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.
