Which of the following technique is used for invitro amplification of gene?

Which one of the following is NOT a technique used for direct transfer of foreign DNA into the protoplast of host cell without using vector?

In tissue culture technique, to maintain aseptic conditions the nutrient medium is autoclaved at constant pressure of _________ $\mathrm{lb} / \mathrm{sq}$ inch for continuous __________ minutes.

$$ \text { Match the restriction enzyme in Column I with their recognition sequences in Column II. } $$