Question

Besides a high voltage shock, what is another method to make E. coli competent to take up "naked" DNA?

a. high concentrations of calcium ions followed by high temperature
b. high concentrations of calcium ions and several hours on ice
c. large amounts of DNA added directly to a bacterial culture growing at 37C
d. high concentrations of minerals followed by high temperature
e. a high voltage shock is the only way to make E. coli competent

Answer 1

E. coli cells can be made competent to take up DNA by incubating with calcium ions and then applying heat shock, which allows DNA to enter the cells through pores formed in the membrane. So the correct option is a.

Step-by-step explanation:

Besides a high voltage shock, another method to make E. coli competent to take up DNA is by using high concentrations of calcium ions followed by a heat shock. This process involves incubating E. coli cells with an ice-cold solution of 50 mM calcium chloride at 4°C for 30 minutes, which allows the DNA molecules to attach to the cell exterior. Following this, the cells are briefly incubated at 42°C, which causes the formation of pores in the membrane, allowing DNA to enter the cell.

Answer 2

The correct answer is a. high concentrations of calcium ions followed by high temperature.

Step-by-step explanation:

Apart from high voltage shock, calcium chloride heat shock method can be used to make E. coli competent to take up naked DNA. As both DNA and lipids contain phosphorus and phosphorus contains negative charge, therefore, ice-chilled CaCl₂ makes cation bridge between phosphorylated lipid of cell membrane and phosphorus present in the DNA.

This allows the naked DNA to attach to the cell surface of bacteria. Then cells are given heat shock which creates gaps(holes) in the membrane and makes the cells competent to take up the naked DNA by these holes.

Then the transformed cells can be selected by screening methods like growing transformed cells on media containing a suitable antibiotics.