Scientific investigation is an iterative process, for which reproducibility is key. Reproducibility, in turn, requires accuracy and precision—particularly in measurement. The unsung superheroes of accuracy and precision in the research lab are the members of your local Metrology Department. According to Promega Senior Metrologist, Keela Sniadach, it’s good when the metrology department remains unsung and behind the scenes because that means everything is working properly.
Holy Pipettes, Scientists! We have a metrology department?! Wait…what’s metrology again?
Metrology (the scientific study of measurement) got its start in France, when it was proposed that an international length standard be based on a natural source. It was from this start that the International System of Units (SI), the modern metric system of measurement, was born.
Metrology even has its own day: May 20, which is the anniversary of the day that the International Bureau of Weights and Measures (BIPM) was created by the Meter Convention in Paris in 1875. The job of BIPM is to ensure worldwide standards of measurement.
This blog was written by guest blogger and 2018 Promega Social Media Intern Logan Godfrey.
Only 30 years ago, the polymerase chain reaction (PCR)
was used for the first time, allowing the exponential amplification of a specific
DNA segment. A small amount of DNA could now be replicated until there was
enough of it to study accurately, even allowing sequencing of the amplified DNA.
This was a massive breakthrough that produced immediate effects in the fields
of forensics and life science research. Since these technologies were first
introduced however, the molecular biology research laboratory has been the sole
domain of PCR and DNA sequencing.
While an amazing revolution, application of a technology
such as DNA sequencing is limited by the size and cost of DNA sequencers, which
in turn restricts accessibility. However, recent breakthroughs are allowing DNA
sequencing to take place in jungles, the arctic, and even space—giving science
the opportunity to reach further, faster than ever before.
The newfound accessibility of DNA sequencing means a
marriage between fields of science that were previously largely unacquainted.
The disciplines of genomics and wildlife biology/ecology have largely progressed
independently. Wildlife biology is practiced in the field through observations
and macro-level assessments, and genomics, largely, has developed in a lab
setting. Leading the charge in the convergence of wildlife biology and genomics
is Field Projects International.
2017 finds Promega on the road visiting cities all across the United States. This year we are presenting workshops from leaders in the forensics community on topics like maximizing success with challenging samples, improving laboratory efficiency and reducing backlogs, and new tools and technologies for the forensics laboratory. This highly popular workshop series is a great way to learn from your peers about new techniques and workflows and network with other forensics experts in your region.
“In Georgia, the legend says
That you must close your windows
At night to keep it out of the house
The glass is tinged with green, even so
As tendrils crawl over the fields…”
—James Dickey (1)
I grew up in Georgia, where on a hot, humid summer day you could almost hear the hiss of growing vegetation, especially the Kudzu as it climbed over fence posts and encroached upon the roadside, the king of invasive species. In Florida you worry about the alligators along the roadside if you have a flat tire; in Georgia, beware the Kudzu.
Invasive species, animal and plant, are an issue in all ecosytems. Imported from distant (and not-so-distant) areas both by accident and misguided intent, invasive species are species that have escaped the checks and balances of natural competitors and predators that existed in their native habitats. This lack of predation and competition enables them to outcompete and overrun other species.
It can be easy to forget that Promega is a manufacturing business. Hidden within the well-designed walls of the company’s cGMP Feynman Center, as well as in other facilities on the Madison campus, technicians operate hundreds of machines that manufacture, dispense and package Promega reagents day in and day out. Keeping those high-tech machines running at peak performance is critical, requiring immense skill, precision and even artistry. That’s where Promega Machinist Technician Travis Beyer comes in.
“I get to make stuff,” says Travis who is not afraid to show his enthusiasm for his craft while describing the best part of his job. “There’s a product at the end of the day. Plus I get to support science, and make things that support people’s lives. That’s cool.”
I get to make stuff. There’s a product at the end of the day. Plus I get to support science, and make things that support people’s lives. That’s cool.
The da Vinci Center, another artfully designed building on the Madison campus, houses the Promega machine shop where Travis does his work designing or improving on parts for newer manufacturing equipment or reverse engineering broken or worn parts no longer available for older equipment that still serves its purpose. He makes every machine part imaginable from drive shafts to sensor brackets to filling forks, and his work is critical to manufacturing businesses like Promega, where a downed piece of equipment can cause costly production delays.
It began at a sink. Advancing from Dishwasher to Production Manager might seem like an unusual career path, but after speaking with Kris Pearson, the Custom/OEM Production Manager at Promega, it appears perfectly ordinary. I was thrilled to meet with her and discuss both the broad strokes and gritty details of working in Custom/OEM Manufacturing. Continue reading “Careers in Science: Kris Pearson, Custom/OEM Production Manager”
Today’s blog post is written by guest blogger Sarah Kolb, Marketing Coordinator for our North America Branch, and new employee at Promega.
As a new member to the North America Marketing team, I was unsure of what to expect going into my first national sales meeting with Promega, but what I took away from this meeting was incredibly eye opening. The North America Branch Sales meeting is an opportunity to get all of the members of the North American branch together to learn about new products, connect with the different strategic business units about product application and network with each other to learn how to better the lives of our customers. The year’s meeting occurred in May in the Ideation room at Promega Headquarters in Madison, Wisconsin.
The room itself is not your typical conference space. An antique car resides in the space, and you can find art-work from all over world nestled in corners, and on the walls and shelves. All around the room, collections of unique furniture are arranged to stimulate conversation. Ideation created an atmosphere of creativity, community and collaboration, which contributed to the overall success of the meeting.
Ever think about the kinds of challenges R&D scientists run up against in the course of developing a new product? The development of the Maxwell® RSC ccfDNA (circulating cell-free DNA) Plasma Kit is a particularly interesting example. Its path to commercialization was characterized by a number of unexpected technical hurdles, yet each was overcome through creative troubleshooting and aided by valuable collaborations across departments. All had a hand in finally launching the kit last August.
The product’s launch was an exciting milestone for Promega as research interest in the role of ccfDNA as biomarkers in human disease continues to grow. Elevated levels of ccfDNA have now been reported in patients with cancer, inflammatory disease, infections and cardiovascular disease. In pregnant women, up to 10% of ccfDNA can be attributed to the fetus, so critical fetal DNA analysis can now be conducted through maternal blood samples. There are many advantages in the ability to isolate and analyze ccfDNA, so the development of a kit with high throughput capability was a priority for the Nucleic Acid Purification R&D team. Continue reading “The Making of a Promega Product: Teamwork = Success for the Maxwell RSC® ccfDNA Plasma Kit”
The path to drug development is strewn with obstacles: Identifying targets; configuring assays to help identify targets or drugs; uncovering the right compound to affect the selected target without off-target effects and screening multiple compounds to eliminate or identify potential drugs. Without the right tools, compounds or target, identifying potential disease therapies becomes nearly impossible.
When it comes to a drug target for cancer, the Ras protein family is at the top of the list because the proteins are expressed ubiquitiously and found mutated in many types of cancer. Because Ras proteins are involved in transducing signals from the surface of cells, many of the resulting mutations produce an activated Ras, inducing uncontrolled expression of the genes that Ras controls. Ras proteins are small GTPases (20–25kDa) that comprise a larger superfamily of proteins divided into five subfamilies: Ras, Rho, Rab, Arf, and Ran. These proteins control diverse cellular activities, including cellular differentiation, proliferation, cell division, nuclear import and export, and vesicle transport. GTPases are guanosine-nucleotide-binding proteins with affinity for GDP or GTP and are able to hydrolyze GTP. When bound to GTP, GTPases are active (turned on) and interact with downstream proteins in the signaling cascade. When GTPases are bound to GDP, the proteins are inactivated (turned off) and no longer transduce signals. Continue reading “A Better GTPase Assay for Drug Development”
Therapeutic monoclonal antibodies are large, complex molecules that undergo numerous post translational modifications (PTMs). In-depth characterization of antibody PTMs remains a significant hurdle because their large size (~150 kDa) makes mass spectrometry analysis extremely challenging.
IdeS protease specifically cleaves IgGs into Fab and Fc fragments. This enzyme is highly specific and cleaves human IgG specifically at one site in the lower hinge region. Because of the exquisite specificity of the enzyme, it produces highly homogeneous Fc and Fab fragments which are then readily analyzed using techniques such as mass spectrometry or HPLC.
One of the drawbacks of IdeS is that it exhibits poor activity against mouse IgGs. IdeZ Protease is an immunoglobulin-degrading enzyme from Streptococcus equi subspecies zooepidemicus. It is an engineered recombinant protease overexpressed in E. coli. Like IdeS Protease, IdeZ Protease specifically cleaves IgG molecules below the hinge region to yield F(ab′)2 and Fc fragments. Reduction of the digestion products produces three fragments of ~25kDa that are readily analyzed by LC-MS.
One of the key advantages of the IdeZ Protease is that it has significantly improved activity against mouse IgG2a and IgG3 subclasses compared to IdeS Protease. IdeZ Protease does not cleave mouse IgG1 or IgG2b.
Key technical parameters when digesting mouse IgGs utilizing IdeZ are the following:
• Add 1 unit of IdeZ Protease per 1µg of IgG to be digested.
• IdeZ Protease is most active in buffers at or near neutral pH. The recommended digestion buffer is 50mM sodium phosphate, 150mM NaCl (pH 6.6).
• Mouse IgG2a and IgG3 typically require 2–4 hours at 37°C for complete digestion.
• IdeZ Protease has a histidine tag for easy removal if so desired.