prepGEM Universal
Catalog Number: MG-PUN0050, MG-PUN0100, MG-PUN0500, MG-PUN1000
Extracts DNA from blood, saliva, and tissue including animal (ear punches, mouse tails, insect) and cell culture
Produces single-stranded DNA for STR, PCR, qPCR, whole genome amplification, amplicon sequencing
Perfect for CRISPR/Cas9 gene editing with NGS
Each extraction kit contains prepGEM, Histosolv, BLUE buffer, RED+ buffer, and ORANGE+ buffer
prepGEM Universal is MicroGEM’s comprehensive solution for effortless high-yield DNA extraction from blood, saliva, and tissues. It is also perfectly suited for extracting DNA from cell cultures. All within a single tube, this method ensures no loss of DNA, and as a result, it excels with low cell number extractions.
This kit is an excellent tool for researchers performing confirmatory genotyping of transfected cells following CRISPR/Cas9 gene editing.
prepGEM Universal utilizes our thermophilic proteinase by activation with a simple temperature change. The whole process is easily automated in a single tube using standard laboratory equipment. A standard thermocycler can be programmed to carry out the extraction.
- Stream-lined approach provides DNA in approximately 15 minutes
- Inhibitor-free solutions mean no harsh chemical washes or multiple steps are required
- Closed-tube extraction protects the integrity of the sample
- Simplified, hands-off workflow frees up technician time, reduces lab costs, and significantly reduces hazardous/plastic waste
Quick-Start Guide: prepGEM Universal
Cell Culture
MicroGEM’s single-step, closed-tube, temperature-controlled lysis and extraction is a robust solution for loss-less DNA extraction from cultured cells.
When working with cultured cells, it is often necessary to isolate DNA from a large number of samples presented in small volumes. These are not limitations for MicroGEM extractions.
When performing gene editing via CRISPR/Cas9, genotyping with Next Gen Sequencing (NGS) is required to confirm the target gene has been successfully modified. This task requires working with very few cells. It is laborious, expensive and difficult with common purification techniques
prepGEM Universal gives linear results from 200,000 cells down to a single cell. Working volumes can be scaled to sub-microlitre volumes making it perfect for working with low cell numbers. Speed, yield and integrity make prepGEM Universal the ideal nucleic acid extraction method for researchers working with cell cultures for gene editing and expression.
Many universities, research hospitals, government entities, and private companies, have already adopted prepGEM Universal into their workflow. The single-step, closed-tube temperature-controlled lysis and extraction is highly desirable because MicroGEM extraction buffers are compatible with downstream reactions. Primers and PCR reagents can be added directly to the extract with no further purification.
For RNA extraction from cultured cells, please see our RNAGEM product.
CRISPR/ CAS
Zhang, Y., Schmid, B., Nielsen, T. T., Nielsen, J. E., Clausen, C., Hyttel, P., . . . Freude, K. K. (2016). Generation of a human induced pluripotent stem cell line via CRISPR-Cas9 mediated integration of a site-specific homozygous mutation in CHMP2B. Stem Cell Research,17(1), 151-153. doi:10.1016/j.scr.2016.06.005. (Genotyping) (pdf)
Yang, L., Yang, J. L., Byrne, S., Pan, J., & Church, G. M. (2014). CRISPR/Cas9-Directed Genome Editing of Cultured Cells. Current Protocols in Molecular Biology. doi:10.1002/0471142727.mb3101s107. (Protocol 3) (pdf)
Mouse or Rat Tails
Biomedical research frequently uses rodents as animal models. In many cases, precise genotypes of the animals are necessary, but often this step is time-consuming.
With the number of animals required to generate data, reagent and labor costs can become prohibitive. This problem can be solved with efficient, low-cost DNA extraction.
PDQeX prepGEM Universal is an automated solution for producing quality DNAs in a low-cost automated system. The DNA is ideal for PCR and qPCR and, with qPCR/HRM, can be used for sexing the animal far more reliably than judging the anogenial distance.
Application Note: Rapid, reliable DNA prep for mouse genotyping using prepGEM (pdf)
Blood - Life Sciences
prepGEM Universal leverages our thermophilic proteinase which is activated with a simple temperature change. The whole process is easily automated in a single tube using standard laboratory equipment. A standard thermocycler can be programmed to carry out the extraction.
Example Results Liquid Blood Extractions: 5 µl of whole blood from an EDTA BD Vacutainer® and fresh blood from a finger-prick was extracted using the PDQeX prepGEM kit. 5 µl of the extract (total volume 100 µl) was used in a 20 µl qPCR reaction. The plots of the two samples are shown below.
Insect
Insects eat between 5-20% of the most important grain crops produced around the world. Further, insect-related vector-borne diseases account for more than 17% of all infectious diseases, causing more than 700,000 deaths annually and millions of cases of dengue, malaria, trypanosomiasis, Lyme disease and many more. Much of this destruction and disease is preventable with timely interventions and prevention strategies.
There are many reasons why it is important to identify an insect at the species level such as guiding insecticide control measures, identifying quarantine pests, and distinguishing disease vector insects. These types of assessments need to be conducted with high levels of accuracy while maintaining high rates of throughput.
The DNA extraction process is often complicated when the sample material is very small and difficult to manipulate such as the legs of insects, larvae and/or eggs. Historically, extracting DNA from a small sample may result in tremendous variations in yield and quality.
MicroGEM's prepGEM Universal reagent kit provides rapid and simple single-tube extractions of single-stranded DNA from intact or crushed insects and insect parts, either fresh or frozen. The single-tube method enables extraction from small samples, even one insect, and ensures all of the nucleic acid in the sample is preserved.
The following publications provide examples of research enabled by prepGEM for nucleic acid extractions from a variety of insect samples:
Agriculture
Studies show that prepGEM is ideal for detection of pest organisms and for the characterization of potential biological control agents.
Natural enemies of armored scales (Hemiptera: Diaspididae) and soft scales (Hemiptera: Coccidae) in Chile: Molecular and morphological identification
https://doi.org/10.1371/journal.pone.0205475
Investigating Biological Control Agents for Controlling Invasive Populations of the Mealybug Pseudococcus comstocki in France
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0157965
Detection of spotted-wing drosophila, Drosophila suzukii
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0098934
Detection of Pacific fruit fly, Bactrocera xanthodes
https://www.cambridge.org/core/journals/bulletin-of-entomological-research/article/dna-barcoding-and-realtime-pcr-detection-of-bactrocera-xanthodes-tephritidae-diptera-complex/82BF63629E12C3DEA22BAB5A7149FC46
Detection of African fig fly, Zaprionus indianus
http://www.bulletinofinsectology.org/pdfarticles/vol70-2017-057-062kremmer.pdf
Phylogenetic analysis of pest South Pacific sap beetle species
https://www.ncbi.nlm.nih.gov/pubmed/22613670
Characterising Aphelinid wasps as a biological control agent
https://link.springer.com/article/10.1007/s10526-016-9752-1
Characterising a Nematode species as a biological control agent
https://www.nzpps.org/journal/65/nzpp_651610.pdf
Identification of scale insects on kiwifruit in New Zealand
https://microgembio.com/wp-content/uploads/2019/10/MAPN005_20191002_Insect_prepGEM.pdf
Disease Vector Insects
These studies demonstrate that prepGEM extractions are suitable for research identifying and monitoring disease vector insects.
Identification of clade types of malaria vector mosquito in north-eastern Tanzania
https://onlinelibrary.wiley.com/doi/full/10.1111/mve.12274
Monitoring insecticide resistance in malaria vector mosquito in south-east Tanzania
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5782413/
Understanding the population structure of a species of tsetse fly in Uganda
https://parasitesandvectors.biomedcentral.com/articles/10.1186/1756-3305-5-222#Sec2
Laboratory Research
A commonly used model organism is the nematode, C. elegans. prepGEM has been used to study populations of C. elegans with a focus on genetic diversity and reproduction.
Characterising male genotypes of C. elegans in response to a novel environment
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0035811#s3
Understanding genotype distribution during the experimental evolution of C. elegans
https://onlinelibrary.wiley.com/doi/full/10.1111/j.1558-5646.2012.01744.x
Aquaculture
prepGEM has been used in aquaculture to extract DNA from larval rock lobster to determine the species of the sampled organisms. It has also been used to develop a low-cost extraction of DNA from shrimp for genotyping in a breeding program.
DNA barcoding of rock lobster species for feeding studies
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0042757#abstract0
Developing a low-cost method for genotyping shrimp for breeding https://www.sciencedirect.com/science/article/abs/pii/S004484861400297X
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