DNA Purification from Plants: Not All Methods are Equal

pestle and mortar with leavesIsolating DNA from plant tissues is difficult for many reasons.  Unlike animal cells, plant cells have rigid cell walls, often made of tough fibrous material, and contain proteins and enzymes and other compounds such as polysaccharides and polyphenols that play a role in different cellular processes. These compounds can interfere with DNA isolation as well as downstream applications such as PCR.  For these reasons, DNA isolation methods that are used successfully for other sample types may not work well to isolate DNA from plant material. Continue reading

Promega Art Showcases Explore Creativity, Science and the Unknown

A visitor studies a piece at the current Promega Art Showcase.

A visitor studies a piece at the current Promega Art Showcase.

Albert Einstein once wrote: “to raise new questions, new possibilities, to regard old problems from a new angle requires creative imagination and marks real advances in science.” The marriage of science and creativity, it seems, is indispensable for exploration and the discovery of new ideas.

As a life sciences company, Promega supports the work of scientists who are tasked with unraveling mysteries and who ask questions in an effort to get answers that improve the lives of others. Because creative thinking plays a key role in the scientific discovery process, Promega supports artistic exploration in many forms. As an organization, we appreciate that creativity reinforces the type of imagination that inspires scientific progress and innovation.

Since 1996, Promega Corporation has sponsored quarterly art showcases at the Promega BioPharmaceutical Technology Center on East Cheryl Parkway in Fitchburg, WI. This artistic initiative came about in an effort to explore the depths of creativity and science, and to demystify biotechnology and the work of Promega for our community. Promega Art Showcases, which occur four times per year and are open to the public, have featured the work of local, national and international artists, as well as the art of Promega employees. Continue reading

Bringing to a Close Women’s History Month with Some Women in Science

Curiosity.In the United States, the month of March is Women’s History month. So, to bring the month of March to a close, I thought I would highlight some women in science. The National Science Foundation, at Science 360, has some great multimedia science resources, including several profiles of women in science, ranging from graduate students, just making their way in science to established scientists who have made major contributions to their fields of study. The videos highlight ambassadors of science, trying to develop a scientifically literate public and laboratory researchers. I picked two to share with you today. But visit Science 360 to see some others.

The first one I thought I would share is of marine biology, PhD student, Ayana Johnson.

And here is a profile of analytical chemist Mary Wirth from Purdue University:

There are many wonderful profiles of scientists, male and female, at the Science 360 web site, and the fascinating work that they pursue. You can also find out more about the people who do science by visiting the This is What a Scientist Looks Like tumbler. It hasn’t been updated since February 2014, but the profiles that are there are quite nice.

Spring Break Staycation

A picnic and play in a local park can be part of your staycation

A picnic and play in a local park can be part of your staycation

Spring Fever! It seems everyone has it around here. Last week, people were the happiest I have seen in a while, and I’m certain the nicer weather had a lot to do with that. With school-age children, it seems that Spring Break is a hot topic of conversation, but an exotic vacation is not always possible for families. So, what can you do if you aren’t taking a Spring Break trip but still want to do something fun with the family? We’ve done staycations a couple times in the past. An overnight in a motel is like a vacation to my children, so that’s popular, but we are looking for new ideas. Here are a few that have been suggested:

  • Camping out! The weather isn’t quite there yet in Wisconsin, but getting out the sleeping bags and having flashlights in the family room is super fun. No electronics, so you get to spend some time together without movies, cell phones or video games.
  • Movie Marathon! We’ve picked a certain type of movie (robots, monsters, etc.), and made popcorn with sweet treats. Pajamas all day on the couch.
  • Scavenger Hunt! Who doesn’t love one? Take the time to create a really fun one that will take a while to get through. This will keep your family entertained for a couple hours if you do it right.
  • Spend the day like a tourist in your own town! There are so many places that get overlooked in our own communities because we are too busy living our lives to explore. I’ve never been to a local museum here! Do some things that people travel to your city to do and see.
  • Family Board Game Tournament! You will be surprised how much fun you can have playing board games. Mix it up so you have a variety. Prizes and snacks too!

What are your staycation ideas?

Optimizing Trypsin Digestion Parameters for Plasma Proteins

TrypsinLysC Page 2Biomarkers in biological fluids in particular have the potential to inform regarding risk of disease or to allow early detection for more effective treatment. Plasma/serum is considered the universal source of biomarkers. This fluid is, indeed, easily collected, and the important point is that plasma collects proteins from each and every tissue, compared to other fluids such as urine or cerebrospinal fluid. Optimizing experimental conditions (i.e., use of trypsin for the digestion of target proteins) used to discover or monitor biomarkers in plasma is critical to successful detection of biomarkers.

In a recent publication by Proc et al., plasma denaturation/digestion protocols were compared using quanititation methods. In this reference 14 combinations of heat, solvent (acetonitrile, methanol, trifluoroethanol), chaotropic agents (guanidine hydrochloride, urea) and surfactants (sodium dodecyl sulfate (SDS)and sodium deoxycholate (DOC) with effectiveness in improving tryptic digestion. Digestion efficiency was monitored by quantitating the peptides from 45 moderate- to high-abundance plasma proteins using tandem mass spectrometry in multiple reaction mode with a mixture of stable isotope labeled analogues of these peptides as internal standards. In the results, Proc et al. noted that use of either DOC and SDS produced an increase in the overall yield of tryptic peptides. Since SDS is not compatible with mass spectrometry and DOC can be easily remove by acid precipitation, the overall recommendation was the use of DOC with a nine hour digestion procedure.

Literature Cited:

Proc, J.L. et al. (2010) A Quantitative Study of the Effects of Chaotropic  Agents Surfactants and Solvents on the Digestion Efficiency of Human Plasma Proteins by Trypsin J. Proteome Res. 9, 5422–37.

Immune Checkpoint Inhibitors: Has Cancer Met its Match?

The cover of S. Mukerjee's book, The Emporer of All Maladies: The Biology of Cancer. Used courtesy of Wikimedia and WLU.

The cover of S. Mukerjee’s book, The Emporer of All Maladies: The Biology of Cancer. Used courtesy of Wikimedia and WLU.

Dr. Drew M. Pardoll, Johns Hopkins University School of Medicine in Baltimore, in his 2012 review, “The blockade of immune checkpoints in cancer immunotherapy” published in Nature Reviews Cancer (1) writes:

“The myriad of genetic and epigenetic alterations that are characteristic of all cancers provide a diverse set of antigens that the immune system can use to distinguish tumour cells from their normal counterparts.”

Tumors have antigens, so we should be able to address/attack these antigens with our immune system, right?

Various immune mediators as therapeutic agents against cancer have entered and mostly flopped in clinical trials over the past 30 or more years. As a graduate student in the 1980s I remember IL-2 and interferon raising many hopes. More recently, drugs against chronic myeloid leukemia and CLL have shown early promise. However, so far cancer cells have mostly won against these therapies. Yet recent news points to some exciting new therapeutic agents, that over the past 15 years or so, and in and out of clinical trials, are getting a leg up in the cancer battle. These drugs are immune checkpoint inhibitors. Continue reading

Get Ready to Celebrate Pi!

26562342_lWhat will you be doing on 3/14/15 at precisely 9:26:53? Twice the clocks will align with the first few digits of my favorite irrational and transcendental number (3.141592653…) or π. Over 1 trillion digits to the right of the decimal point have been calculated, and they still go on, never making a pattern, never repeating.

Take any circle, that blueberry pie that your grandmother baked would be a good choice. Measure the circumference and divide by the diameter and you will have Pi, the mathematical constant that represents the ratio of circumference to diameter in a circle. You probably encountered Pi in your early mathematics education when learning the formula for the area of a circle (2πr2).

In celebration of Pi, here is a few Pi diversions to brighten your day. Continue reading

Uncovering Protein Autoinhibition Using NanoBRET™ Technology

13305818-protein ligandIntroducing new assays or technologies is meant to make it easier for you to perform research and craft experiments to test hypotheses. However, scientists are creative people, and new technologies or assays may just be the catalyst for a crucial experiment or new data you are seeking. In the case of a recent Proceedings of the National Academy of Sciences USA article, Wang et al. used the principle of our NanoBRET™ assay to understand how ERK1/2 phosphorylation of Rabin8, a guanine nucleotide exchange factor, influenced its configuration and subsequent activation of Rab8, a protein that regulates exocytosis. Continue reading

Neuroscience Explains Harry Potter’s Appeal

BookWithGlassesCurling up with a good book is one of life’s greatest pleasures, whether you’re reading on a tropical beach while on vacation or nestled into your favorite chair at home. As your eyes skim over the words, your mind conjures up images of the events unfolding on the page. Books can take us to fantastic places, real and imaginary, that we will never visit in our lifetime. And while there is some pleasure to be gained from nonfictional books, my favorite books all seem to fall in the realm of fiction. I am not alone. The science fiction and fantasy genre of literature continues to be one of the most popular. Why do so many readers find these types of books so enticing and engaging?

It all comes down to science, specifically neuroscience.

Continue reading

Reduce, Reuse and Recycle: Foam Coolers

12447811_IMG_3195Life science enzymes, cells and reagents are often temperature sensitive, and you need products that arrive cold and ready to work. This means that packaging often requires dry ice, gel ice and foam coolers—challenges for maintaining a small carbon footprint and environmentally responsible shipping and packaging program.

In the last few years, we have moved to unbleached shipping boxes, started using sustainably harvested materials and biodegradable and recyclable air pouches to offer product protection while minimizing negative environmental impact. Continue reading