Monarch Kits for Plasmid Purification
The Monarch Plasmid Miniprep Kit is a rapid and reliable method for the purification of high quality plasmid DNA. Elution in as little as 30 μl provides concentrated DNA for use in downstream applications, such as restriction digests, DNA sequencing, PCR and other enzymatic manipulations.
Protocols are fast and user friendly, saving you valuable time. Reagents include color indicators at certain steps to easily monitor completion.
Video Protocol Monarch Plasmid Miniprep
Designed with sustainability in mind
Columns and bottles have thinner walls, reducing total plastic usage without affecting performance.
Wir erreichen dies durch den Einsatz besonders dünnwandiger Säulchen und Flaschen. So reduzieren wir den Einsatz von Kunststoff und damit das Müllaufkommen im Labor. Kit boxes, inserts, and protocol cards are made from recycled paper.
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Unique Column Design
The design of Monarch miniprep columns eliminates buffer retention and salt carryover, resulting in highly pure plasmid DNA for your downstream needs. Elution can be done in as little as 30 µl, and convenient tab and frosted surfaces facilitate handling and labeling.
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.
Isolate high-quality, concentrated plasmid DNA
Monarch Plasmid Miniprep Kits consistently produce more concentrated plasmid DNA with equivalent yield, purity and functionality as compared to the leading supplier
Preps were performed according to recommended protocols using 1.5 ml aliquots of the same overnight culture. One microliter of each prep was digested with HindIII-HF (NEB #R3104) to linearize the vector and the digests were resolved on a 1% w/v agarose gel.
Learn more about the performance of the Monarch Miniprep Kit on neb.com
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: Plasmid Miniprep Tips
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.
Available Monarch Plasmid Miniprep products:
| PRODUCT | Prod. # | SIZE | INFO | |
|---|---|---|---|---|
| Monarch Plasmid Miniprep Kit* | T1010S/L | 50 / 250 preps |  |
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| Monarch Plasmid Resuspension Buffer (B1) | T1011L | 55 ml | |
|
| Monarch Plasmid Lysis Buffer (B2) | T1012L | 2×27 ml | |
|
| Monarch Plasmid Neutralization Buffer (B3) | T1013L | 110 ml | |
|
| Monarch Plasmid Wash Buffer 1 | T1014L | 2×27 ml | |
|
| Monarch Plasmid Wash Buffer 2 | T1015L | 30 ml | |
|
| Monarch DNA Elution Buffer | T1016L | 25 ml | |
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| Monarch Plasmid Miniprep Columns | T1017L | 100 preps | |
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As of: 04.01.2024
*Permanent low prices – no further discounts apply. Offer closes 31.12.2024.
Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.
