DNABook™ users can extract DNA using either PCR or transformation in E.coli.
Cut out the piece of paper from the DNABook™. It´s recommended that a commercial leather-punch tool with a 2mm diameter punch for cutting the DNA sheet. Alternatively, cut a 2mm x 2mm square from the DNA sheet around the required DNA spot, using an either a sterile scalpel or scissors.
Collecting cDNA inserts using PCR.
Any cDNA inserts can be amplified and collected using 2 PCR techniques, as described below:
Recommended primer sequences: Fwd primer: 5′-TGTAAAACGACGGCCAGT-3′ Rev primer: 5′-AGCGGATAACAATTTCACACAGGA-3′
94 oC 2mins 94 °C 1min } 60 °C 1min } x29 cycles 68 °C 1min 15secs 74 °C 15mins
94 °C 2mins 94 °C 1min } 60 °C 1min } x29 cycles 68 °C 1min 15secs 74 °C 15mins
Transformation Clone isolation by transformation of plasmids obtained from the DNA sheet.