A rapid and low-cost genomic DNA extraction protocol for plant tissue

Mankesh Kumar, Ashutosh Kumar, Deepak Rajpurohit, Priyanka Kumari, Nitish Kumar & Rabiya Parveen

Genomic DNA extraction is the first step in almost every plant molecular biology technique. This includes routine PCR genotyping and marker-assisted breeding. Long-established methods rely on CTAB or phenol-chloroform extraction. Both give high DNA purity, but both are slow. Both also depend on hazardous organic solvents. It uses a sodium dodecyl sulfate (SDS) based extraction buffer, called the Rapid Buffer. A sodium borate (SB) buffer is used alongside it for agarose gel electrophoresis. We give full recipes for both buffers. We also give a complete fifteen-step extraction procedure, with the reasoning behind each step. The article covers agarose gel preparation, supporting laboratory calculations, a comparison with conventional methods and practical guidance on troubleshooting and safety. The protocol avoids phenol and chloroform entirely. It uses small reaction volumes, so many samples can be processed in parallel. The full procedure takes well under an hour of active bench time. This makes it well suited to routine genotyping, high-throughput screening and teaching laboratories.