This blog was written by guest writers Paraj Mandrekar (Technical Services Scientist 3) and Michelle Mandrekar, (Research Scientist 4).
Here are some designer’s notes comparing the Maxwell® RSC Blood DNA and the Maxwell® RSC simplyRNA kit chemistries for nucleic acid extraction.
The Maxwell RSC Blood DNA Kit and Maxwell RSC simplyRNA Blood Kit were both developed from the same non-silica-based purification chemistry and use the same underlying paramagnetic particle. This chemistry is characterized by an extreme binding capacity (the capacity of nucleic acid that can be bound on the particle), leading to both chemistries being capable of isolating large amounts of nucleic acid volumes and then eluting into relatively small volumes (50 µL). It is not unusual with either chemistry to have isolates that exceed 100 ng/µL. Although the chemistries have several similarities, there are some important distinctions between how the two chemistries were designed that influence which kit you choose for your nucleic acid extraction.
Traditionally, scientists have relied on flat,
two-dimensional cell cultures grown on substrates such as tissue culture
polystyrene (TCPS) to study cellular physiology. These models are simple and
cost-effective to culture and process. Within the last decade, however, three-dimensional
(3D) cell cultures have become increasingly popular because they are more
physiologically relevant and better represent in vivo conditions.
Isolating 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 “DNA Purification from Plants: Not All Methods are Equal”
Back in the dark ages, when I was a graduate student, my idea of “automated” plasmid DNA extraction involved performing home-brew, “toothpick preps” in “strip tubes” or , if I was really feeling ambitious, a 96-well plate.
I would get just enough DNA to check for the presence of an insert, but the quality of the DNA was too low and the quantity too small to even consider using it for any other downstream experiments like amplification.
And increased throughput for other nucleic acid extraction needs? Nope. If I wanted genomic DNA, RNA or high-quality plasmid DNA, I spent time with columns and tubes, giving each sample my undivided individual attention.
Remember cesium chloride preps for RNA isolation? Even with the advent of column purification, which greatly simplified and standardized my protocols, nucleic acid purification was still a manual task that required a lot of time and effort to get the high-quality product I needed.
Doing the experiments that would answer the questions that I really wanted to ask (those “downstream experiments”), meant spending time at the bench performing careful (if tedious) work to isolate and clean up the highest quality nucleic acid possible. Even then inconsistency in sample prep could wreak havoc on downstream work.
Fortunately, for the modern scientist, personal, bench top automation, has progressed far beyond the toothpick and the strip tube to quality-tested, reliable nucleic acid extraction platforms like the Maxwell® Rapid Sample Concentrator (RSC).
The Maxwell® RSC improves sample preparation consistency, eliminating variability associated with manual handling, and your downstream results will reflect this consistency. With the RSC you can extract DNA or RNA from up to 16 samples in approximately 1 hour and viral total nucleic acids in less than an hour.
The instrument is easy to use: simply load the sample, push a button and walk away. Cross contamination is minimized and the instrument is supported by the Promega technical support and service you have come to trust over the past 35 years.
Want to know more about how the Maxwell® RSC can give you the freedom to focus on the work that interests you the most? To learn more, click here.
Guest Post from Promega Technical Services Scientist, Caroline Davis.
On a snowy day in January, someone stole the cookies that were to be served with lunch from the Rome Corners Intermediate School cafeteria. The kids were distraught. What should they do? Luckily, the Green 2 Team science class was there with Promega’s Technical Service Outreach team (and Paraj Mandrekar, Senior Research Scientist and Green 2 Team Dad) to help.
The students realized that the thief had taken a bite out of a strawberry and left part of it behind, along with his DNA. After a short discussion on what DNA is and why you would want to isolate DNA, the 6th graders extracted DNA from strawberries using household reagents under the guidance of the Promega scientists. The students used pipettors, beakers, microfuge tubes and flipper racks, giving the students a glimpse of the tools that scientists use everyday in in a molecular biology lab. Continue reading “Who Stole the Cookies? Technical Services Scientists Offer DNA Labs to Area Schools”
Yesterday 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”
In honor of Halloween, here is a Top Ten Uses of Pumpkin list for your enjoyment:
10. Means of transportation on the high seas or emergency flotation device (pumpkin boats; see the video).
9. Elementary Math Lessons. Determine circumference and radius. How much does a pumpkin weigh? Estimate the number of seeds. Check out pumpkin math ideas here.
8. Cholesterol-lowering snack. After ruthlessly scooping out the innards of a pumpkin, clean the seeds, bake them in the oven and enjoy your healthy snack. Pumpkin seeds contain phytosterols, compounds that that have been shown to reduce levels of LDL cholesterol, as well as magnesium, a mineral needed in the diet.
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