6533b7d8fe1ef96bd126acaf
RESEARCH PRODUCT
A Rapid Method for Amplification of Plastome DNA-Fragments from Spinacia oleracea by PCR
Helmut ReiländerHelmut TrippAloysius Wildsubject
Reaction conditionsSpinaciabiologyPhysiologyPlant Sciencebiology.organism_classificationMolecular biologylaw.inventionChloroplastchemistry.chemical_compoundchemistryChloroplast DNAlawOrganelleChenopodiaceaeAgronomy and Crop ScienceDNAPolymerase chain reactiondescription
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 method described one can amplify any known plastome-fragment in a few hours.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1993-07-01 | Journal of Plant Physiology |