Halloween Costumes: Retro Science Style

Your Promega Connections bloggers were sitting around reminiscing the other day, “Back when I was in the lab…”. Kind of like Thanksgiving Dinner among your elderly relatives, it wasn’t long before we were one-upping each other with horror stories from our days at the bench–stories that included escape artist rats, a leaky sequencing gel apparatus, and the iconic radioactively contaminated post doc.

We decided to turn that conversation, with a lot of help from our favorite science cartoonist Ed Himelblau, into retro Halloween costumes based on our memories of things we used to do in the lab that don’t seem like such a great idea now. Enjoy…and if you have a few retro horror science costume ideas of your own you would like to add, feel free to comment.

First up: Cesium Chloride Preps.

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From Napkin Sketch to “Custom Kit”: CloneWeaver® Workflow Builder Gets Your Cloning Organized

20161018_150403Let’s face it, most lab techs and purchasing agents aren’t all that happy when you send them an Instagram picture of your latest lunchroom-napkin cloning strategy as your order form for your next big cloning experiment. So we have created the CloneWeaver® Workflow Builder. You can transfer your brilliance easily from that lunchroom napkin to an orderly email or print out of every vector, enzyme, purification kit, and transfection reagent your next big molecular cloning experiment requires. You can even save your one-of-a-kind “cloning kit” for future endeavors.

The CloneWeaver® tool will walk you through every step of the molecular cloning process from selecting a vector to finding a transfection reagent for mammalian cells. So if you are starting a new project, we are with you every step of the way. We will help you find restriction enzymes and even remind you about markers and biochemicals that you may want to have on hand for your experiment. Within the tool we have links to additional resources like our RE Tool and catalog pages if you need more help.

clone_weaverAlready have a favorite vector and a freezer full of restriction enzymes? No problem, skip those steps and move on to getting the perfectly sized nucleic acid markers or the particular polymerase your experiment requires.

Are you teaching a molecular genetics course? CloneWeaver® workflow builder is perfect for creating the list of laboratory reagents you are going to need for your students—and you will have this same list as a starting point for other lab experiments or classes later on because you can save the lists that you build. You can even pass them along to other professors.

So, if molecular cloning is in your future, let us help you get organized. Try the CloneWeaver® Workflow Builder.

Pack a Little Science into Your Summer with Advanced Courses from BTCI

This new cGMP facility was built to have the least effect on the natural environment as possible--surrounded by rain gardens and native plantings, using grey water for flushing toilets--sustainable growth was the goal.

Summer on the Prairie at Promega–Study science surrounded by birds, bees, flowers and amazing prairie.

Summer, a much-looked forward to season. We typically pack in the activities and make the most of the daylight. We work hard and we play hard.  This summer will be no exception, and at the BTC Institute, we are already getting set to host as many students as we can. We will see middle and high schoolers, K-12 teachers, college students, graduate students, college and university faculty and staff, and professionals in the biotech community under our roof at some point. You may want to join us too!

Our programs for advanced learners, geared toward the graduate student or biotech professional, offer much more than just a rigorous immersion in molecular biology theory and practice. Held at the BTC Institute at Promega Headquarters, they are taught by highly knowledgeable scientists, coming from both industry and academia. These instructors offer a wealth of information and share their expertise as well as life experiences with students. Informal discussions about career trajectories and access to industry are important added benefits to attending these off-campus workshops. Continue reading

Welcome to Your Biotechnology Field Trip at the BTC Institute!

Kaukauna High School students arrive at the BTC for a biotechnology fieldtrip.

Kaukauna High School students arrive at the BTC for a biotechnology fieldtrip.

BTCI provides our students an opportunity that they could never get in the classroom.
—Jim Geoffrey, Biology Teacher, Kaukauna High School

Your bus has arrived and parked in the circular driveway at the front of the BioPharmaceutical Technology Center on the Promega Corporation campus in Fitchburg, WI. Your BTC Institute hosts – and instructors – for your field trip are Barbara Bielec (K-12 Program Director) and Ryan Olson (Biotechnology Instructor). They’ll greet you in the Atrium and direct you to a conference room where you can leave coats and backpacks, and then to the lab you’ll be working in during your visit.

Here’s a taste of what happened next for students from Random Lake High School and Wonewoc High School on December 3rd, and from Kaukauna High School on December 4th. Continue reading

Tips for Successful Dual-Reporter Assays

Dual-Reporter-AssayRecently, one of my fellow bloggers described some of the advantages of using dual-reporter assays (including our Dual-Luciferase®, Dual-Glo® Luciferase and our new NanoDLR™ assay debuting soon). These assays are relatively easy to understand in principle. Use a primary and secondary reporter vector transiently transfected into your favorite mammalian cell line. The primary reporter is commonly used as a marker for a gene, promoter, or response element of interest. The secondary reporter drives a steady level of expression of a different marker. We can use that second marker to normalize the changes in expression of the primary under the assumption that the secondary marker is unaffected by what is being experimentally manipulated.

While there are many advantages to dual-reporter assays, they require careful planning to avoid common pitfalls. Here’s what you can do to avoid repeating some of the common mistakes we see with new users: Continue reading

Reflections on Summer 2014 Courses

Group Photo from the 2014 Core Techniques in Protein and Genetic Engineering Course Held at the BTC Institute July 14-18. Photo credit: BTCI

Group Photo from the 2014 Core Techniques in Protein and Genetic Engineering Course Held at the BTC Institute July 14-18. Photo credit: BTCI

One of the things that I encourage all of the students I interact with in BTC Institute courses to do in order to boost retention and make meaning out of the activities that we do in class is to reflect on their experiences. Reflection is one way to connect new knowledge to past experience and get it to really stick in the brain, among other things.

Taking my own advice to heart, I use this space to ponder some interesting aspects of these experiences from my own perspective. This summer, I worked with 65 students and over 25 instructors to deliver four weeks of intensive instruction in molecular biology applied to a wide range of research areas. Continue reading

Elegant Experiments that Changed the World

Crystallographic structure of HIV reversed transcriptase. Wikimedia Commons

Crystallographic structure of HIV reverse transcriptase. Wikimedia Commons

Today, reverse transcriptases are commonplace molecular biology tools, easy to obtain and routinely used in labs for everyday cloning and gene expression analysis experiments. Reverse transcriptase inhibitors have also found widespread use as antiviral drugs in the treatment of retroviral infections.

It’s easy to forget that the existence of reverse transcriptase activity—the ability to convert an RNA template into DNA—was once a revolutionary notion not easily accepted by the scientific community. The idea that RNA could be the template for DNA synthesis challenged the “DNA–>RNA–> Protein” central dogma of molecular biology.

The foundational studies that proved the existence of a reverse transcriptase activity in RNA tumor viruses were described in two papers published back-to-back in Nature in June, 1970. Two of the authors of these studies, Howard Temin of the University of Wisconsin and David Baltimore of the Massachusetts Institute of Technology, were awarded a Nobel Prize for their work in 1975.

In appreciation of the significance of these papers, the editorial introduction published in Nature at the time states:

This discovery, if upheld, will have important implications not only for carcinogenesis by RNA viruses but also for the general understanding of genetic transcription: apparently the classical process of information transfer from DNA to RNA can be inverted.

Before these papers were published, it was known that successful infection of cells by RNA tumor viruses required DNA synthesis. Formation of virions could be inhibited by Actinomycin D—an inhibitor of DNA-dependent RNA polymerase—so it was known that synthesis of viral RNA from a DNA template was part of the viral life cycle. The existence of an intracellular DNA viral genome was therefore indicated, and had been postulated by Temin in the mid 1960’s. However, proof of the mechanism whereby this DNA template was generated from the RNA genome of the infecting virus remained elusive. Continue reading

Why You Don’t Need to Select a Wavelength for a Luciferase Assay

4621CAIt’s a question I’m asked probably once a week. “What wavelength do I select on my luminometer when performing a luciferase assay?” The question is a good and not altogether unexpected one, especially for those unfamiliar or new to bioluminescent assays. The answer is that in most cases, you don’t and in fact shouldn’t select a wavelength (the exception to this rule is if you’re measuring light emitted in two simultaneous luciferase reactions). To understand why requires a bit of an explanation of absorbance, fluorescence, and luminescence assays, and the differences among them.

Absorbance, fluorescence, and luminescence assays are all means to quantify something of interest, be that a genetic reporter, cell viability, cytotoxicity, apoptosis, or other markers. In principle, they are all similar. For example, a genetic reporter assay is an indicator of gene expression. The promoter of a gene of interest can be cloned upstream of a reporter such as β-galactosidase, GFP, or firefly luciferase. The amount of each of these reporters that is transcribed into mRNA and translated into protein by the cell is indicative of the endogenous expression of the gene of interest. Continue reading

ProK: An Old ‘Pro’ That is Still In The Game

Proteinase K Ribbon Structure ImageSource=RCSB PDB; StructureID=4b5l; DOI=http://dx.doi.org/10.2210/pdb4b5l/pdb;

Proteinase K Ribbon Structure ImageSource=RCSB PDB; StructureID=4b5l; DOI=http://dx.doi.org/10.2210/pdb4b5l/pdb;

If you enter any molecular lab asking to borrow some Proteinase K, lab members are likely to answer: “I know we have it. Let me see where it is”. Sometimes the enzyme will be found to have expired. The lab may also have struggled with power outages or freezer malfunctions in the past. But the lab still decides to keep the enzyme. One may rightly ask – why do labs hang on to Proteinase K even when it has been stored under sub-standard conditions? Continue reading

DNA Purification, Quantitation and Analysis Explained

WebinarsYesterday I listened in on the Webinar “Getting the Most Out of Your DNA Analysis from Purification to Downstream Assays”, presented by Eric Vincent–a Product Manager in the Promega Genomics group.

This is the webinar for you if you have ever wondered about the relative advantages and disadvantages of the many methods available for DNA purification, quantitation and analysis, or if you are comparing options for low- to high-throughput DNA purification. Eric presents a clear analyses of each of the steps in a basic DNA workflow: Purification, Quantitation, Quality Determination, and Downstream Analysis, providing key considerations and detailing the potential limitations of the methods commonly used at each step.

The DNA purification method chosen has an affect on the quality and integrity of the DNA isolated, and can therefore affect performance in downstream assays. Accuracy of quantitation also affects success, and the various downstream assays themselves (such as end-point PCR, qPCR, and sequencing) each have different sensitivities to factors such as DNA yield, quality, and integrity, and the presence of inhibitors. Continue reading