Protocols, Primers, and other Particulars - Barrington Lab
PCR Protocol (for rbcL and trnL-F)
The chloroplast gene rbcL and the chloroplast spacer between the trnL and trnF genes (hereafter trnL-F) were amplified using the polymerase chain reaction (PCR) in 25 mL aliquots with the following reaction components: 50-150 ng of genomic DNA, 1X Ex Taq buffer (TaKaRa, Madison, Wisconsin, USA), 200 mmol/L of each dNTP, 0.1 mmol/L of each primer, and 0.625 U Ex Taq (TaKaRa) using a Techne thermal cycler (model TC-312) with thermal cycling conditions as follows: for rbcL initial denaturation was for 5 min at 95°C, followed by 40 cycles of 1 min at 94°C, 1 min at 50°C, 2 min at 72°C, finishing with a final extension of 72°C for 8 min; for trnL-F initial denaturation was 5 min at 95°C, followed by 40 cycles of 1 min at 94°C, 1 min at 50°C, 2 min at 72°C, finishing with 72°C for 8 min.
List of Accessions on hand in the Barrington lab (with thanks to all who have contributed.)