ZyGEM has developed an integrated pipeline that takes you from sampling leaves in an orchard, greenhouse or the field through to DNA preparation.
The sampling step is simple, robust and reliable. Using the ZyGEM Crusher tool, leaf samples are processed onto barcoded storage cards. The sample collection takes less than a minute.
From this card, 2-3 punches are placed in the PDQeX system and DNA is eluted in 15 minutes. The system is capable of processing 24 samples in a single 15 minute run, although smaller batch numbers can be used without wasting reagents or plasticware.
This workflow simplifies the process of collecting, archiving and extracting nucleic acids from leaves, seeds, roots, stems, cotyledons, as well as other succulent sample types such as insects.
Using ZyGEM's PDQeX technology for DNA extraction, total processing time is approximately 15 minutes from a plant sample to PCR-ready DNA.
The phytoGEM collection cards allow for plant samples to be stored at room temperature for months at a time, removing the need for freezing of samples and cold storage.
As the PDQeX machine is fully portable, collection and extraction can be done in the greenhouse with no need to take samples back to the lab.
Downstream applications such as qPCR, minION and Ion Torrent NGS have all been performed on PDQeX processed samples.
An example of using PDQeX extractions in the field can be seen in this video.
PDQeX 2400
The PDQeX 2400 is a rapid, robust DNA extraction device that can achieve PCR and Human profile-ready DNA in under ten minutes. The compact size means it can easily be placed in a laminar flow hood or on the benchtop.
PDQeX System
The PDQeX 2400 is a robust DNA extraction system that prepares PCR-ready DNA within 7-20 minutes or less. Its small footprint (200 x 200 mm), disposable extractor cartridges and 24 sample capacity make it perfectly suited to the needs of all types of laboratories, protecting the integrity of the sample and the safety of lab personnel.
Sample types that can be extracted using the PDQeX include:
- Blood
- Animal tissue
- Plant tissue
- Saliva
- Swabs
- Cell Culture
- Bacteria
- Semen
- Insects
- Nematodes
The EA1 protease was selected from a panel of over 100 proteases in the ZyGEM culture collection for a number of valuable characteristics that benefited the process of nucleic acid extraction.
The enzyme has a narrow window of actiivty at the relatively high temperature of 75°C, which means it is effectively 'frozen' at ambient temperature, and inactivation is achieved readily above 90°C.
The high temperature of actiivity enhances the proteolytic activity on mesophilic substrates, allowing the use of the protease, to extract DNA or RNA, in PCR and RT-PCR compatible buffers. No chaotropes, salts or pushing the pH up to 10.7! This liberates the researcher from the need for solid phase purification in many applications. It also allows for the detection of lower abundance targets, as the extraction is lossless.
The narrow window of thermal activity allows the EA1 protease to be used in conjunction with mesophilic enzymes, in a single tube extraction. This is illustrated with the prepGEM Bacteria kit, in which lysozyme and the prepGEM EA1 protease are sequentially activated and removed/inactivated with incubations at 37°C, 75°C and 95°C.
When applied to the extraction of total nucleic acid from small cell populations, the EA1 protease - in the RNAGEM formulation - offers unparalled performance by taking advantage of these key characteristics. The high temperature of activity ensures ready cellular degradation, RNAse and DNAse degradation and, importantly, the removal of protein material from the RNA templates released, ensuring maximum reverse transcriptase processivity.
There are a number of papers and app notes published using the ZyGEM technology, from single cryptosporidium to buccal swabs for forensic profiling. Some of these are in our downloads section, but the peer review material is best found here
RNAGEM Tissue and RNAGEM Tissue plus offer extraordinary power in the analysis of small cell populations or plate based cell culture assays.
Delivering linear yields of DNA and RNA from 1-200,000 cells, these kits allow the immediate gene expression profiling of FACS sorted primary and cultured cells, and laser captured samples.
When used in 96 or 384 well format, these kits dramatically reduce the cost of RNA preparation in cell culture applications, and provide greater sensitivity when profiling low abundance transcripts.
For more information on RNAGEM, please click here, for application notes please visit the download section.
- The ZyGEM protease rapidly degrades RNAse A and other RNA-degrading enzymes (see QC Page)
- Closed–tube (protects sample integrity, minimises cross-contamination and improves bio-safety)
- Hands–off (saves labour costs and allows for high throughput batch processing)
- One-step (reduces the opportunity for processing to alter the biological composition of the sample)
- Easily automated (compatible with most low cost, off-the-shelf liquid handling robots)
- Optimal for small sample sizes and low cell numbers (with linear yields from 10 – 100,000 cells)
- Can be adapted for single-cell work
- Fast and easy. Compare the workflows of RNAGEM and Trizol
ZyGEM offers an expanding range of DNA and RNA extraction kits, defined by the process of Temperature Controlled Extraction, or the use of heat to activate sequential enzymatic steps to control the extraction process through to the heat inactivation of the reagents.
The kits are scaleable, automatable and lossless, and yield DNA suitable for most PCR based applications including routine PCR for cloning or sequencing, qPCR, STR-PCR, WGA and SNP detection using platforms such as the MassArray and other PCR based detection methods. The scalability allows the same reagents to be used on tissue samples from mouse tails to laser capture microdissected tissue, with linear yields of DNA release from 1 to 200,000 cells.
The ZyGEM reagents, rely on the use of a highly active, thermostable protease, that allows the extraction of DNA in buffers that are highly compatible with downstream PCR and RT-PCR. This allows for more lysate to be carried into the downstream PCR, offering greater sensitivity in detecting low abundance DNA species in mixed samples. For more information on the EA1 protease click here