The Monarch PCR & DNA Cleanup Kit rapidly and reliably purifies up to 5 μg of concentrated, high-quality DNA from PCR and other enzymatic reactions. The kit utilizes a bind/wash/elute workflow with minimal incubation and spin times. The columns ensure zero buffer retention and no carryover of contaminants, enabling elution of sample in volumes as low as 6 μl. The buffers provided have been optimized, and do not require monitoring of pH. Eluted DNA is ready for use in restriction digests, DNA sequencing, ligation and other enzymatic manipulations. Designed with sustainability in mind, Monarch kits use significantly less plastic and responsibly-sourced, recyclable packaging. The protocol can also be modified to enable the purification of smaller DNA fragments, including oligonucleotides and ssDNA.View our videos on protocols, tips, and recycling Monarch.
APPLICATIONS
PCR cleanup
DNA from PCR reactions can be purified after amplification to remove polymerases, primers, detergents, dNTPs, etc.
Enzymatic reaction cleanup
Restriction enzymes and modifying enzymes such as ligases, kinases, nucleases, phosphatases are efficiently removed, allowing for effective desalting and concentration of the DNA sample.
cDNA cleanup
DNA/RNA complexes can be purified post-reverse transcription/amplification to enable removal of the RT and polymerase as well as nucleotides.
Labeling cleanup
Unincorporated radiolabeled or fluorescently labeled nucleotides can be removed from the DNA substrate
Plasmid cleanup
Plasmid preps from unknown sources may contain inhibitors and unwanted contaminants. Purification and concentration can be easily achieved using this kit.
Oligonucleotide cleanup
ssDNA oligonucleotides (≥ 18 nt) and dsDNA fragments (≥15 bp) can be purified using the Oligonucleotide Cleanup Protocol.
Monarch DNA Cleanup Column DesignMonarch columns are designed for performanceMonarch columns are designed without a frit, which eliminates buffer retention and the risk of carryover contamination, providing fast, worry-free DNA purification.Specifications
DNA Sample Type:
DNA from PCR and other enzymatic reactions (e.g., restriction digests, kinase reactions, ligations).ssDNA or dsDNA oligonucleotides from enzymatic reactions can also be purified using the Oligonucleotide Cleanup Protocol.
Binding Capacity:
up to 5 μg
DNA Size Range:
~50 bp to 25 kbDNA≥ 15 bp to 25 kb (dsDNA) and DNA≥ 18 nt to 10 kb (ssDNA) can also be purified using the Oligonucleotide Cleanup Protocol.
Typical Recovery:
DNA (50 bp to 10 kb): 70–90% DNA (11–23 kb): 50–70%ssDNA≥ 18 nt and dsDNA≥ 15 bp: 70-85%
Elution Volume:
≥ 6 μl
Purity:
A260/280 > 1.8 and A260/230 > 1.8
Protocol Time:
5 minutes of spin and incubation time
Compatible DownstreamApplications:
ligation, restriction digestion, labeling and other enzymaticmanipulations, library construction and DNA sequencing.
Monarch PCR & DNA Cleanup Kit (5 µg) ProtocolMonarch PCR & DNA Cleanup Kit (5 μg) performs equivalently to the leading supplierPreps were performed according to recommended protocols. 1 μg of a 3 kb DNA fragment was incubated with 1 μM primers and OneTaq® Quick-Load® 2X Master Mix (NEB #M0486). DNA was eluted in 20 μl (NEB) and 40 μl (Qiagen) Elution Buffer. Half of the total elution volume was digested with 5 units of DraIII-HF® (NEB #R3510). The digest and the unused portion of the elution were resolved on a 1% w/v agarose gel along with a representative sample of the starting material.Monarch PCR & DNA Cleanup Kit (5 μg) removes low molecular weight primers from dsDNA samplesThree independent amplicons (267 bp, 520 bp, 1003 bp) were spiked with two oligonucleotides (16-mer, 24-mer) to a final concentration of 1 μM. Half of each mix was purified with the Monarch PCR & DNA Cleanup Kit (5 μg) following the included protocol. Equivalent fractions of the original mixture and the eluted material were resolved on a 20% TBE acrylamide gel at 100V for one hour and stained with SYBR Green II.
This product is related to the following categories:
DNA Cleanup Products,
Nucleic Acid Purification Products
This product can be used in the following applications: