Full name: 
Dr. Tytti Vanhala

Affiliation

Swedish University of Agricultural Sciences
Research interests: 

Genetics

Content published on MolMeth

Lepidium campestre has proven to be one of the more difficult plant species for DNA isolation. Here we describe a method that has proven to produce good and clean DNA for RAD sequencing purposes. There are several stages to the protocol. First the leaf samples are pulverized and proteins are broken down. Second stage is the actual extraction of DNA which should follow directly after the pulverization and protein breakdown. Third stage comprises of quality control and RNA removal. This is essential if the samples are intended for example RAD sequencing. The fourth and final stage of the process is to estimate the DNA concentration. This is not as straightforward as it may sound, because L. campestre appears to have long strands of genomic DNA coiling and clumping together. If this type of clumping and uneven distribution of DNA happens in the solution, for example Nanodrop will not provide accurate estimates of DNA concentrations. We describe a way to relax the DNA and make it possible to obtain more accurate estimates of DNA concentrations.
Lepidium campestre has proven to be one of the more difficult plant species for DNA isolation. Here we describe a method that has proven to produce good and clean DNA for RAD sequencing purposes. There are several stages to the protocol. First the leaf samples are pulverized and proteins are broken down. Second stage is the actual extraction of DNA which should follow directly after the pulverization and protein breakdown. Third stage comprises of quality control and RNA removal. This is essential if the samples are intended for example RAD sequencing. The fourth and final stage of the process is to estimate the DNA concentration. This is not as straightforward as it may sound, because L. campestre appears to have long strands of genomic DNA coiling and clumping together. If this type of clumping and uneven distribution of DNA happens in the solution, for example Nanodrop will not provide accurate estimates of DNA concentrations. We describe a way to relax the DNA and make it possible to obtain more accurate estimates of DNA concentrations.