2. Calculate Transformation Efficiency of the following experiment using the information and the hypothetical results listed below: -3 . PGLO concentration: 6X10 ug/ul Cells incubated with 560 ul CaCl2 transformation solution and split into two tubes (-PGLO and + PGLO). 20 ul PGLO solution to the tube of + PGLO 200 ul LB 50 ul spread on agar plate 213 colonies of transformants on LB/Amp plate
Answers
Answer:
Nucleic acids do not enter the cells under their own power; they require assistance in crossing
physical barriers at the cell boundary and in reaching an intracellular site where they can be
expressed or replicated.
The standard method of making the bacterial cell permeable to DNA involves treatment of
bacteria with calcium ions and giving electrical current or heat shock treatment to the cells.
Chemically induced competence followed by transformation is a commonly used technique
to introduce plasmids or other DNA fragments into the bacteria. On the contrast, during
electroporation, the exposure of cells to an electrical discharge reversibly destabilizes their
membranes and transiently induces the formation of aqueous pathways or membrane pores
TRANSFORMATION
that potentiate the entry of DNA molecules. This method of electroporation is comparatively
rapid, simple and efficient.
Explanation: