Isolating gDNA from Plant samples
The field of plant genomics is one of the fastest evolving fields in life sciences today. With this development, there is an increased need for high quality genomic DNA from leave and seed samples. This series of articles describes the changes noticed within the field of plant genomics, together with a number of challenges and solutions. The information and data in these articles is obtained during the optimization and validation of one of our projects at one of the leading plant breeding companies.
This first article will describe the demands and needs within plant genomics today. The challenges during upscaling will be described in the second article and this series will be finalized with a description of the solutions provided by GC biotech.
Evolutions in Plant Genomics
The introduction of novel techniques such as Next Generation Sequencing results in an increasing amount of samples. In parallel the demand for a higher quality of the isolated DNA is another result of the introduction of these techniques. As the amount of samples increases, the need for an efficient process with a focus on cost reduction is one of the highest priorities within the field.
Within the field of Plant genomics, there are several types of projects. Some projects focus on a limited variety of crops, while other projects focus on a really wide variety of crops.
When the variety of crops is small, the process of isolation is easily adapted per crop. However, when the amount of samples increases, the need for automated processes increases as well. With a wide variety of crops, the adaptations needed per crop will complicate the use of automated processes.
Isolating DNA from plant material with magnetic beads
To be able to isolate DNA from material such as seeds and leaves from plants, grinding balls are added to the sample. Most often these are ceramic or tungsten. When placed into a tissue grinder, the samples are grinded into a fine powder.
The process of DNA isolation from plant material starts with a lysis step. The most common processes all use a detergent in the lysis buffer resulting in a high level of foam formation if lysis buffer is added during the grinding step.
If the sample preparation has been performed, the process continues with a binding step and several wash buffers. As the wash buffers contain alcohol, the process also contains a drying step. After alcohol removal, the isolated gDNA is eluted from the beads with an elution buffer. The eluted DNA is transferred into a clean plate and can be used for downstream applications.
The next post
This article describes the evolution of DNA isolation in the field of Plant genomics. The next article will go into detail on the challenges as seen today. The need for high quality gDNA and the high throughput.