lab tips

Reaching Out for Lab Research Experience

Posted on by Promega

Today’s guest blog is written by Melissa Martin, a global marketing intern with Promega this summer. She will be a senior this fall at the University of Wisconsin-Madison where she is double majoring in zoology and life sciences communication, with a certificate in environmental studies.

Congrats! You are attending a university and pursuing a challenging, yet rewarding, undergraduate science degree. Getting to this moment probably included lots of late nights spent studying or worrying while applying to your dream college. However, now that you are here you will find that classes provide a lot of information. You can even take your education one step further by getting hands-on experience in a research lab.

Working in a lab is not only about making your resume look good. It offers a real-world experience that directly enhances your learning experience and can even guide your future. For example, your experiences in the lab can teach you basic skills (pipetting, determining concentrations, performing titrations, etc.) that will be useful in a variety of science professions.

Continue reading “Reaching Out for Lab Research Experience”

Choosing Your Subcloning Strategy

Posted on by Kelly Grooms

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.

Top Ten Tips for Successful PCR

Posted on by Kelly Grooms

We decided to revisit a popular blog from our Promega Connections past for those of you in the amplification world. Enjoy:

magnesium-31

    • Modify reaction buffer composition to adjust pH and salt concentration.
    • Titrate the amount of DNA polymerase.
    • Add PCR enhancers such as BSA, betaine, DMSO, nonionic detergents, formamide or (NH4)2SO4.
    • Switch to hot-start PCR.
    • Optimize cycle number and cycling parameters, including denaturation and extension times.
    • Choose PCR primer sequences wisely.
    • Determine optimal DNA template quantity.
    • Clean up your DNA template to remove PCR inhibitors.
    • Determine the optimal annealing temperature of your PCR primer pair.

[Drum roll please]…and the  most important thing you can do to improve your PCR results is:

  • Titrate the magnesium concentration.

The Price for Convenience May Not Be That Pricey After All

Posted on by admin

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?

DNA Purification, Quantitation and Analysis Explained

Posted on by Isobel Maciver

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 “DNA Purification, Quantitation and Analysis Explained”