Figure 2

DNA amplification using PMOS. Panel A contains the design of EO's and TO's that combine to form the forward primers for amplification of the ftsZ gene. Vertical bars indicate clamp regions of hybridisation between the EO and TO. Capital letters show actual sequence of the oligonucleotides and small, underlined letters indicate the extended region of the EO. N represents a position with either A, T, G or C. S represents a position with either G or C nucleotide. Panel B contains the DNA template sequence of E. coli ftsZ and includes the recognition sequence for the M13 reverse primer (double underlined) within the plasmid pFC1. The position of the forward primers (formed by extended EO's) is indicated by the dotted and solid underlining. Panel C contains a photograph of a 1% agarose gel produced after amplification of the ftsZ sequence. Lane 1 contained a DNA molecular weight marker with corresponding sizes shown on the left. Lane 2 contains an amplification reaction with oligonucleotides EC10 and M13 reverse (positive control). Lane 3 contains an amplification reaction with oligonucleotides E128, T128 and M13 reverse. Lane 4 contains an amplification reaction with oligonucleotides E128 and M13 reverse (negative control). Lane 5 contains an amplification reaction with oligonucleotides E382, T382 and M13 reverse. Lane 6 contains an amplification reaction with oligonucleotides E382 and M13 reverse (negative control).