A Rapid Method for Amplification of Plastome DNA-Fragments from Spinacia oleracea by PCR

Summary Polymerase Chain Reaction (PCR) was used as a very powerful technique to amplify DNA-fragments. Isolated chloroplasts and a crude suspension of small leaf pieces of Spinacia oleracea were osmotically shocked and subsequently heated. Aliquots of the resulting homogenate including the organelle DNA were directly added to basic reaction mixtures to carry out PCR without additional isolation- or purification steps. A 180 by DNA-fragment from the psbA-gene was amplified in both cases. In parallel, plasmid-DNA (pHT1) bearing the psbA-gene was subjected to PCR retaining unchanged reaction conditions. Identical DNA-fragments were obtained, as the following sequencing revealed. With the meth…