The Monarch Genomic DNA Purification Kit is a universal kit for DNA extraction and purification from a wide variety of cell types, including blood, cells, tissues and tough-to-lyse samples (including bacteria and yeast). The kit includes lysis buffers for various input materials, Proteinase K for homogenization of some sample types, and RNase A for efficient RNA removal. Genomic DNA (gDNA) is eluted with high yield and purity, and with excellent integrity (high molecular weight), ready for use in downstream applications including qPCR, long range PCR, and NGS library prep (including for long read sequencing platforms).
Reasons to Choose Monarch for Genomic DNA Extraction and Purification
Purify high-quality, genomic DNA from a broad range of sample types (cells, blood, tissues, and more).
The Monarch Genomic DNA Purification Kit provides excellent yields of higher quality, higher molecular weight DNA than the Qiagen® DNeasy® Blood & Tissue Kit
Agilent Technologies® 4200 TapeStation® Genomic DNA ScreenTape was used for analysis of blood, cultured cell, and tissue samples purified using the relevant protocols of the Monarch Genomic DNA Purification Kit and the Qiagen DNeasy Blood & Tissue Kit. gDNA was eluted in 100 μl and 1/100 of the eluates (~1 μl) was loaded on a Genomic DNA ScreenTape. Starting materials used: 100 μl fresh human whole blood, 100 μl frozen pig blood, 1x106 HeLa cells and 10 mg frozen tissue powder. Monarch-purified gDNA samples typically show peak sizes 50 – 70 kb and DINs of ~9. DNeasy-purified gDNA peak sizes are typically <30 kb with DINs ~7-8. DNeasy kits produce lower yields and low A260/A230 ratios for liver, brain, muscle and frozen blood.
Achieve high yields from challenging tissues that are difficult for many commercial kits (e.g. fatty, fibrous, and soft organ tissues).
The Monarch Genomic DNA Purification Kit provides excellent yields with tissues that are problematic for other commercial kits
Duplicate 10 mg samples of RNAlater®-stabilized rat tissue were cut to small pieces and subsequently lysed and purified according to the protocols provided with each kit. Optional RNase A steps were included. Elution was carried out with 100 µl elution buffer provided in the respective kits. Yields displayed are averages of the duplicate samples, and represent the genomic DNA yield after correcting for the RNA content as determined by LC-MS. Results indicate that the Monarch Genomic DNA Purification Kit provides excellent yields for a wide range of tissues.
Prevent co-purification of RNA with our optimized buffers and included RNase A, enabling more accurate quantification.
DNA purified with the Monarch Genomic DNA Purification Kit has significantly lower residual RNA across all sample types
RNA content present in genomic DNA eluates from various kits was evaluated by LC-MS. All samples were processed in duplicate according to manufacturers’ recommendations and were eluted in 100 μl. Starting materials used: 100 μl human blood, 1x106 HeLa cells, 10 mg of RNAlater-stabilized mouse (tail) and rat tissue samples (others). 1 μg of each sample was treated with the Nucleoside Digestion Mix (NEB #M0649) and subjected to LC-MS.
Values displayed are averages of duplicate measures and indicate the percentage of riboguanoside (rG) versus the total amount of ribo- and deoxyriboguanoside in the samples. Actual RNA content may be lower for all samples, since rG is more abundantly co-purified in silica preps than other RNA bases. The Monarch Genomic DNA Purification Kit consistently delivers residual RNA below 1%–2% levels, which is usually undetectable with most analysis methods and lower than what is seen for other commercial kits.
Generate highly-intact genomic DNA suitable for long-range PCR, qPCR, NGS and other downstream applications.
The Monarch Genomic DNA Purification Kit generates high quality genomic DNA 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 and 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 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.
The Monarch Genomic DNA Purification Kit generates excellent input material for NGS library preparation with NEBNext® kits for Illumina®
A. Duplicate libraries were made from 100 ng HeLa cell gDNA purified with Monarch (orange) or Qiagen DNeasy Mini Kit (blue) using the NEBNext Ultra II FS DNA Library Prep Kit for Illumina (NEB #E7805). Libraries were sequenced on an Illumina MiSeq. Reads were mapped using Bowtie 2.2.4 and GC coverage was calculated using Picard’s CollectGCBiasMetrics (v1.117). Expected normalized coverage of 1.0 is indicated by the horizontal grey line, the number of 100 bp regions at each %GC is indicated by the vertical grey bars, and the colored lines represent the normalized coverage for each library. Monarch GC coverage matched Qiagen DNeasy results.
B. High yield libraries are achieved from Monarch-purified gDNA. Library yields of the samples described above were assessed on an Agilent Technologies® 2100 BioAnalyzer using a High Sensitivity DNA Kit.
Monarch Genomic DNA Kit generates high quality DNA for Nanopore sequencing
HeLa cell genomic DNA was extracted using either the Monarch Genomic DNA Purification Kit or the Qiagen DNeasy Blood & Tissue Kit. One microgram of purified DNA was used to prepare Oxford Nanopore Technology (ONT) sequencing libraries following the ONT 1D Ligation Sequencing Kit (SQK-LSK109) protocol without DNA fragmentation. Libraries were loaded on a GridION (Flow cell R9.4.1) and the data was collected for 48 hrs. Libraries produced using the Monarch gDNA exceeded the Qiagen libraries on common sequencing metrics including: A. total sequencing data collected, B. read quality, and C. read length. Data was generated using NanoComp (Bioinformatics, Volume 34, Issue 15, 1 August 2018, Pages 2666–2669).
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