Curiosity Unleashed: Wisconsin Science Festival 2014

Our buggy conversation starter at the Wisconsin Science Festival

Our buggy conversation starter at the Wisconsin Science Festival

“Oh wait Mom, look, bugs!”

And she was off. The next thing I knew she had a pale greenish-blue tobacco hornworm caterpillar in her hand.

“I’ve never seen anything like this before.” Nor had I.

She turned it upside down and started tickling its legs with a finger of her free hand.

“What does it eat? What will it turn into? How big will it get?” Rapid fire questions at the student who was manning the table of bugs. “Ooo cool. Look Mom he has a stick bug on his shirt. What does a woolly bear caterpillar become? What’s the name of that beetle? Where does it live? Why is that hornworm so much more active than the one I’m holding?”

We both took a really good look at the beating heart just underneath the dorsal skin of the very active hornworm that was about to pupate.

That was one adventure. There were many more.

Promega was one of many sponsors of this year’s Wisconsin Science Festival: Curiosity Unleashed 2014. Continue reading

Tips for a Social Media User

Social%20Networks_smallerAs an HR professional, I attend different seminars and conferences to obtain credits for my HR certification. We had a SHRM (Society for Human Resources Management) state conference last week, and I learned all kinds of new strategies involving recruitment, succession planning, employee engagement and change management. One topic was present in every session I was in: social media. How to recruit through social media, engage employees in social media, and how to maintain your company and personal brand through social media.

One session in particular was focused solely on social media, and how it affects the processes of hiring, discipline and discharge. Continue reading

Extraction of Plant DNA Made Easy

By Trillium1946 at en.wikipedia (Transferred from en.wikipedia) [Public domain], from Wikimedia Commons

My one attempt at working with plant DNA when I was at the lab bench was trying to create a shotgun library from a rice BAC. Never have I needed to isolate nucleic acid from the source material, but based on my conversations with plant scientists, it can be problematic endeavor between the tough tissue and the compounds that can copurify during extraction and inhibit downstream applications. And if you want to isolate DNA or DNA from plant samples in an automated format, that just adds to the difficulties. Here I review an Applications in Plant Sciences article that compares DNA isolation using the Maxwell® 16 System with two other methods on 25 different plant species samples. The authors note that Promega provided the Maxwell® 16 instrument, DNA isolation cartridges and advice on its use. Continue reading

Choosing Your Subcloning Strategy

Before you begin your subcloning, you need to know: The restriction enzyme (RE) sites available for subcloning in your parent vector multiple cloning region (or in the insert if you need to digest the insert); the RE sites available in the destination vector multiple cloning region (MCR); and if these same sites also occur in your insert. Once you know this information, you can use the chart below to decide which subcloning strategy to use.

4498MA-[Converted]

To learn more about subcloning, visit our Subcloning Notebook.

Use of Cell-Free Technology to Evaluate Nuclease (TALEN) Activity on Target DNA

ImageSource=RCSB PDB; StructureID=1qpf; DOI=http://dx.doi.org/10.2210/pdb1qpf/pdb;

ImageSource=RCSB PDB; StructureID=1qpf; DOI=http://dx.doi.org/10.2210/pdb1qpf/pdb;

Transcriptional activator-like effector nucleases (TALENs) have rapidly become a technique of choice for precision genome engineering. TALENs are custom-designed nucleases that consist of a modular DNA-binding domain fused to a monomeric, C-terminal FokI nuclease domain (1). TALENs work in pairs and are designed to recognize and bind to tandem-oriented sequences in genomic DNA, separated by a short spacer (15–30 bp). TALEN binding causes dimerization and activation of the FokI nuclease domains, which results in cleavage of the DNA within the spacer region. Small insertions or deletions (indels) are frequently introduced at this site, as the result of errors made during DNA repair by nonhomologous end-joining (NHEJ). These indels can be up to several hundred base pairs in length and result in frameshift mutations that lead to the production of truncated or nonfunctional proteins.

Successful use of TALENs for inducing targeted mutations has been reported in many conventional models, for example: mice, Xenopus and D. melanogaster. TALENs are also reported to be functional in a variety of other invertebrate arthropods, including mosquitos,silkworm and cricket. A recent publication (2) illustrates the use of TALEN technology for the genetic manipulation in P. dumerilii (marine ragworm). Continue reading

Highlights from ISHI25

ISHI25 logoLast week (September 29–October 2), I was one of almost 1,000 people who attended the 25th International Symposium on Human Identification (ISHI25) in Phoenix, Arizona. This scientific meeting brings together DNA analysts from forensic and paternity labs, research scientists and others with an interest in DNA-based identification to learn about new technologies, policy and process changes, and current and future trends in DNA typing. There were so many great presentations and learning opportunities, how do I pick just a few of them to highlight?

Continue reading

Free Webinar: Protein Reference Materials for Mass Spectrometry

MSextractcroppedOnce the domain of analytical chemistry labs, mass spectrometry instruments are now used in basic life science research, drug discovery research, environmental and industrial laboratories, and in many other biological laboratories. These instruments, which are used to perform critical analyses, need to be monitored for performance and quality.

How do you determine the system suitability and quality for your experiments? How do you evaluate a sample preparation protocol for mass spec analysis to ensure you get the best results possible without introducing artifacts? And, if you are trying to optimize the instrument method, what standard do you use ?

Currently, there is no commercially available reference reagent to help you monitor and test all LC (liquid chromatography) and MS (mass spec) parameters, particularly sensitivity and dynamic range, in a single run.

But what if there were an optimized peptide mixture that could report all key instrument performance parameters in a single run? Even better, what if that mixture came with free software to make analysis of all of the data you can generate even easier?

The upcoming free webinar: The 6 × 5 LC-MS/MS Peptide Reference Mixture and Software Analysis Tool describes such a standard reagent and analysis software.
If you are interested in learning about standardized reagent and software to help you monitor your instrument or optimize methods or protocols, register for this free webinar today!

Detecting Inhibition of Protein Interactions in vivo

Protein Interactions with NanoBRETIn a paper published in the September issue of ACS Medicinal Chemistry Letters, researchers from GlaxoSmithKline in the UK and Germany report on the discovery, binding mode and structure:activity relationship of a new, potent BRPF1 (bromodomain and PHD finger containing protein family) inhibitor. This paper came to our attention as it is one of the first publications to apply Promega NanoBRET technology in an vivo assay that reversibly measures the interaction of protein partners. The technology enabled the identification of a novel inhibitor compound that disrupts the chromatin binding of this relatively unstudied class of bromodomain proteins.

What exactly are bromodomains and why do they matter?
Bromodomains are regions (~100 amino acids) within chromatin regulator proteins that recognize and “read” acetylated lysine residues on histones. These acetylated lysines act as docking stations for regulatory protein complexes via binding of the bromodomain region. Because of their role in chromatin binding and gene regulation, bromodomains have attracted interest as potential targets for anti-cancer treatments. Although some bromodomain-containing proteins (e.g., those in the bromodomain and extraterminal domain (BET) subfamily) are well characterized and have been identified as potential therapeutic targets, others are less well understood. Continue reading

Opportunities for High School Students to Learn at the BTC Institute

Paul Simon famously sang about what it was like to engage as a learner in a high school environment—though his lack of education certainly hasn’t hurt him any, I do wonder about reading the “writing on the wall”. Frequently, in Education, we talk about the challenges of preparing students for careers that have yet to be invented. What to do?

One major initiative within K-16 education can broadly be referred to as “21st Century Skills”—those that are needed for individuals to be successful contributors in a society where concrete goals are moving targets. Though we don’t know the exact details, we’re pretty sure that there are some basic elements that all people will need to be successful contributors to society.

Partnership for 21st Century Skills has built a framework for understanding and aligning our education system toward these skills:

Photocredit: Partnership for 21st Century Skills http://www.p21.org/about-us/p21-framework

Photocredit: Partnership for 21st Century Skills http://www.p21.org/about-us/p21-framework

Applying this model to educational programming takes a lot of innovation and hard work on the part of instructors as well as students. However, students who have the opportunity to engage with a teaching and learning system that makes use of these concepts can reap big rewards when it comes to being able to understand how their learning can be applied to solving problems. Here at the BTC Institute, we have been fortunate to work with the Dane County (Wisconsin) School Consortium to develop two offerings for high school students in the area of biotechnology that really work within this model and give students the contextualization they need to develop academic and career skills. Continue reading

Riboprobes: RNA Probes Are Still Valuable Research Tools

9613ca[1]Riboprobes are RNA probes that can be produced by in vitro transcription of cloned DNA inserted in a suitable plasmid downstream of a viral promoter.
Viruses code for their own RNA polymerases, which are highly specific for the viral promoters. Using these enzymes, labeled NTPs, and inserts in both forward and reverse orientations, both sense and antisense riboprobes can be generated from a cloned gene.
Transcription of RNA is performed with the appropriate RNA polymerase (T3, T7 or SP6), depending on the RNA polymerase promoter sites present in the chosen vector. Because these polymerases are extremely promoter-specific (i.e., there is almost no transcriptional cross talk), virtually homogeneous RNA can be obtained using plasmid DNA as the template in a transcription reaction. When it is desirable to copy only insert DNA sequences, the plasmid is linearized at an appropriate restriction site before the transcription reaction and only discrete “run-off” transcripts are obtained, virtually free of vector sequences. RNA transcripts may be used to generate radioactive probes for hybridization to Northern and Southern blots, plaque and colony lifts as well as non-radioactive probes (i.e, labeled with digoxgenin)for in situ hybridization.

Recent references using riboprobes include: Continue reading