Figure 1: Test for presence of DNA without DNA purification.
In previous experiments, performing DNA isolation before PCR for the SspC gene failed to return bands of DNA in gel electrophoresis. In procedure, B. subtilis was used directly in the PCR mixture. Lane 1 contained the DNA ladder for bothe gels A and B. In A, lane 2 and in B, lanes 2 and 3 contain the PCR products using unaltered B. sutilis. In lane 1, the DNA ladder is visible. Gel A showed no bands in lane 2. Gel B showed a band in both lanes 2 and 3, present around 300 base pairs.
Figure 2: Sequences of oligonucleotide primers with their corresponding melting temperatures.
The location of the primers in indicated in bold font on the targeted 720 base-pair sequence. The sequence of this 720 base-pair sequence that codes for the SspC gene is underlined. The forward primer is designed to bind to the complementary strand of DNA during PCR and move in the 5¢ to 3¢ direction, replicating the identical sequence as above. The reverse primer is designed to bind to the DNA sequence at the end of the sequence above in an antiparallel manner and move in the 5¢ to 3¢ direction (Kunst et al., 1997).
