As part of our research on the best practices for soil microbial DNA extraction, we collect a wide variety of samples for product development. So when we were developing the protocols for the PowerLyzer 24 Homogenizer 24 Homogenizer, we wanted a protocol that worked for most of the samples tested. Our work on homogenization and bead tubes previously showed that depending on the soil, sometimes a different bead type could give you an increased yield of DNA. We decided to do a similar study using the PowerLyzer 24 Homogenizer to ask the question: what is the difference in DNA yields and integrity using high powered bead beating between two different soils using the same protocols? It is not uncommon for people to simply adopt a protocol from a paper for their soil type without doing any optimization. But, does one protocol really work best for every soil?Read More
HOMOGENIZATION AND BEAD TUBE METHODS PART 2: DETERMINING THE BEST HOMOGENIZATION PROTOCOL FOR ANY SOIL
2/1/2017 3:04 PM
1/25/2017 2:04 PM
One of the most efficient ways to extract nucleic acids from a sample is by smashing it against a hard surface repeatedly under high speed until cell walls and membranes crush from the pressure and release their internal contents. In other words: bead beating.
Bead beating is a great way to do what enzymes take hours to accomplish and sometimes never fully succeed in, which is cell lysis to release DNA or RNA for isolation. While enzymes can be successful for DNA isolation from a limited number of sample types, results are achieved a lot faster if you break down the matrix first. And RNA cannot be isolated in a timely fashion without the use of some kind of mechanical maceration.
The questions inevitably arise though, how hard do I need to beat to lyse my sample and how do I know what bead type to use?Read More