Precision will be reduced, but yield will be increased

Optimisation of a PCR reaction is often a compromise between the competing demands for precision, efficiency and yield. Although the specific effects may vary, generally, increasing the annealing temperature will increase non-specific primer binding and reduce precision. Increasing the length of the elongation phase will reduce the proportion of incomplete newly-synthesised strands and therefore increase yield. In this case, the potential effect on efficiency is unclear. Increasing the elongation phase would increase the reaction time, but the time taken to ramp down to a lower annealing temperature would be reduced.

A restriction enzyme cleaves a circular plasmid vector to produce fragments : 1kb, 0.3kb, 2kb, 1kb and 0.3kb. How many bands will be visible on agarose gel?

The attractive interactions lead to ______energy.

Wild type E. coli metabolizes the sugar lactose by expressing the enzyme ß-galactosidase. You have isolated a mutant that you call lac1–, which cannot synthesize ß-galactosidase and cannot grow on lactose (Lac–). During an condition isolate a mutation that constitutively expresses abnormally high levels of ßgalactosidase, which you designate lacc. Preliminary P1 transduction experiments indicate that lacc  is linked to the Tn5 insertion.  To map lacc  relative to lac1– you set up two reciprocal crosses. In the first cross you grow P1 on a strain that carries the Tn5 insertion and the lac1– mutation. You then use this lysate to infect a lacc mutant and select for Kanr. From 100 Kanr transductants examined, 20 are Lac–, 76 express ß-galactosidase constitutively and 4 show normal ß-galactosidase expression. In the second cross you grow P1 on a strain that carries the Tn5 insertion and the lacc mutation. You then use this lysate to infect a lac1– mutant, and select for Kanr. From 100 Kanr transductants examined, 81 are Lac– and 19  express ß-galactosidase constitutively. So what will be the correct order of  the Tn5 insertion and the lac1– and lacc  mutations. Express any measured distances as cotransduction frequencies.