Cloning Modified Blunt-ended DNA Fragments into T-Vectors

Tailing blunt-ended DNA fragments with TaqDNA Polymerase allows efficient cloning of these fragments into T-Vectors such as the pGEM®-T Vectors. This method also eliminates some of the requirements of conventional blunt-end cloning — Fewer steps, who can argue with that?

Blue/White colony screening helps you pick only the colonies that have your insert.
Blue/White colony screening helps you pick only the colonies that have your insert.

Continue reading “Cloning Modified Blunt-ended DNA Fragments into T-Vectors”

Tick, Tock! The Molecular Basis of Biological Clocks

A long time ago, before the rise of humans, before the first single celled organisms, before the planet even accumulated atmospheric oxygen, Earth was already turning, creating a 24-hour day-night cycle. It’s no surprise, then, that most living things reflect this cycle in their behavior. Certain plants close their leaves at night, others bloom exclusively at certain times of day. Roosters cock-a-doodle-doo every morning, and I’m drowsy by 9:00 pm every night. These behaviors roughly align with the daylight cycles, but internally they are governed by a set of highly conserved molecular circadian rhythms.

Jeffrey Hall, Michael Rosbash and Michael Young were awarded the 2017 Nobel Prize in Physiology/Medicine for their discoveries relating to molecular circadian rhythms. The official statement from the Nobel Committee reads, “…this year’s Nobel laureates isolated a gene that controls the normal daily biological rhythm. They showed that this gene encodes a protein that accumulates in the cell during the night, and is then degraded during the day. [They exposed] the mechanism governing the self-sustaining clockwork inside the cell.” What, then, does this self-sustaining clockwork look like? And how does it affect our daily lives (1)?

Continue reading “Tick, Tock! The Molecular Basis of Biological Clocks”

What Things Are You Thankful for in Science?

What are you thankful for in science?
What are you thankful for in science?
As the social media lead for Promega, I keep my eye on trends in new media. I have personal accounts that I keep mostly to see what other people are doing. I try hangouts, social networking and other things so that I have an idea of developing practices outside of the biotechnology industry. One activity that has been popular over the last couple of years during the month of November in the United States is the Facebook post of “30 days of thanksgiving”.

I wondered what “thanksgiving” looks like to the research scientist. So I asked:

What are the things you are thankful for in science?

The answers have been as varied as the people I talked to ranging from little things like water bath floats to really big things, like the renewal of your research funding or achieving tenure.

Here are some of the answers from my informal inquiries:

“Tube floaties for water baths.”

—E.V., genomics product manager

“I was always thankful for Geiger counters.”

—K. G., science writer

“Thermal cyclers and Taq Polymerase. As an undergrad I watched someone sit with a timer and move their tubes between water baths at 3 different temperatures, opening tubes and adding polymerase at the end of each cycle. Modern PCR is SOOO much easier.”

—M.M., research scientist

“I am thankful for competent cells. I remember preparing the CaCl2 and doing slow centrifugation. Also thankful for serum-compatible transfection, rapid ligations and online journal access (no longer have to traipse over to the university library to get papers photocopied- uuurrrgggghhh).”

—R.D., technical services scientist

“How about T-vectors for cloning? I was no molecular biologist, but could make a T-vector work.”

—K.K., science writer

“I am thankful for open-access journals and the ability to read the full article without an institutional subscription.”

—S.K., science writer

“I am ever so thankful for ONLINE ORDERING! So awesome. Throw in online technical manuals, on-line support tools, on-line calculators – all are awesome!!”

—A.P., director, scientific courses

“I am thankful for automated sequencing- manual sequencing was laborious and hazardous!!!”

—R.G., technical services scientist

Do any of these resonate with you? What are you thankful for as a scientist? Let us know in the comments.

Is This What a Scientist Looks Like?

scientists-at-workI am the mother of a six-year-old girl who loves to get magazines in the mail. For several years my daughter has received an enjoyed popular kids’ science/international culture magazine. The stories are short and simple, and this magazine usually does a good job of presenting factual information in easy-to-digest forms. Each magazine comes with a set of animal cards, which we have diligently collected.

However, the latest issue that came to our mailbox really got me thinking. The final pages featured artwork by the young readers. I love the idea of featuring the work of the readers.  Usually, my daughter loves seeing what other children her age from around the world draw and take pictures of, and sometimes we have some pretty interesting discussions about the work.

This time though we didn’t spend much time talking about the art work. She wasn’t particularly interested, and I wasn’t sure I what I thought. But I may have missed a teachable moment. The theme for the pages was a Halloween-minded “spooky science”, and all of the pictures were of “mad scientists” alone at work doing presumably nefarious things in their laboratories. Of the eight drawings pictured, six of them pictured scientists that were human, and five of the humans were male. All of them were pale-skinned. The sole female scientist, whose lab featured a certificate with the words “monster maker”, was drawn by a girl. The ages of the children submitting the work ranged from 9 to 14. Continue reading “Is This What a Scientist Looks Like?”

Science Confirms What We’ve Always Suspected: Potato Chips are Irresistible

RatI love potato chips. There’s something very satisfying about the crunch of a good chip. The problem with chips, other than the obvious effect they have on my waistline, is that I can’t eat just one. Neither can my husband, who loves to open a bag of potato chips while I’m preparing dinner! To explain the disappearance of the potato chips, we joke that the chip-eating culprit in our house is not my husband but a giant mouse that has developed a taste for salty snacks.

Recent research presented by Tobias Hoch at a meeting of the American Chemical Society shows that not only do rodents love potato chips but that this attraction may not be due solely to the high ratio of fats and carbohydrates, which is one proposed explanation for the “bet you can’t eat just one” phenomenon. There is something else that makes potato chips irresistible. Continue reading “Science Confirms What We’ve Always Suspected: Potato Chips are Irresistible”

The Price for Convenience May Not Be That Pricey After All

Hour glass

I was having a discussion with my mother just the other day about cleaning products (lively topic, I know). She showed me her newest time saver…prediluted bleach. Huh, I thought. I guess that does save a bit of time, but I couldn’t resist telling her that she was paying triple the price for a whole lot of water. She said, without pause, that it was worth it to her to not have to splash fully concentrated bleach around. A convenience worth paying for, in her words.

I don’t know why this struck me as odd. I pay for convenience all the time as I get older. When I started running gels back in college, I wouldn’t have dreamed of buying a precast gel, but several years into my lab life I found myself running more than 15 gels a week, so precast was really a convenient alternative. When I was a grad student, I poured all of my own plates (and most of the plates for older students, too!). Fast forward a few years, and I running upwards of 300 microbial selective cultures per week. The switch to prepoured plates was a no brainer.

When put in the context of what our time is worth, would you rather be thawing and mixing loading dyes, buffers, stains, reagents, etc., or are you better of grabbing a premixed, room-temp stable dye or ladder/loading dye mix off the shelf and getting on with your research? I think most scientists would agree that these small conveniences allow you to free up a little more time to do the important work you should be doing.

I’m curious…what time savers or convenience items do you find that make your day a little easier in the lab?

Antibiotics and Honey–An Old Solution for a New Problem

Antibiotic DiskThis month saw the publication of a UK Department of Health report on the growing problem of antimicrobial resistance, which included the shocking recommendation that antimicrobial resistance be added to the UK government list of threats to national security alongside terrorism and pandemic flu. In this report, Professor Dame Sally Davies, Chief Medical Officer for the UK, focused on the increasing problem of multidrug resistant organisms–raising the profile of an important issue that many scientists and health-care professionals have warned us about before. A March 12 Nature editorial welcomed the recommendations as a sign that policy makers in the UK are taking the threat of antimicrobial resistance seriously and are prepared to take more steps to address the problem of multidrug-resistant organisms. Continue reading “Antibiotics and Honey–An Old Solution for a New Problem”

Promega Connections: The Year in Review

Your Promega Connections bloggers had a great time bringing you cool science stories, technical tips and assorted other reading material this year, and we want to say a big “Thank you!”  to all of our readers for your time, your comments, and your reblogs.

Here are some of the highlights from 2012.

I can doodle people!
I can doodle people!
In January Kelly blogged about The Making of a Science App, describing the work to create the Cell Signaling iPad app that we released last year. An update is forthcoming early in 2013 (GPCRs!), and we continue to improve our iPad, iPhone and Android apps as well as our web tools for scientists. If you haven’t played with them, check them out. They are all free. Cats, dogs and their humans have found Kari’s blog post about catnip intriguing as well. Continue reading “Promega Connections: The Year in Review”