Nondestructive DNA Extraction Techniques for Soybean (Glycine Max) Seeds

While maintaining high germination rates, genotyping single seeds are useful in breeding and genetics. Nondestructive single-seed genomic DNA extraction protocols using 12 mg cotyledon tissue with a modified cetyl trimethyl ammonium bromide (CTAB) technique and a commercial seed DNA extraction kit using 1 mg cotyledon tissue were developed for dry soybean seeds and cross-verified with leaf DNA analysis. The DNA extracted by these methods was sufficient for polymerase chain reaction (PCR), and the results were reproducible for several endogenous genes and transgenes in D-myo-inositol 3-phosphate synthase (MIPS) mutant seeds (GM-lpa-TW1). This single-seed DNA-analysis method is amenable to rapid high-throughput genotype screening and verifying the genetic purity of seed stocks.