by Ain Nuha Hazlan

1. Obtaining the Gene

ainxxx — 2014-04-14 04:33:50 UTC

1) Breaking open the cells through chemical & physical methods
2) Purify the DNA using phenol and proteinases, this is to remove proteins
3) Cutting the genes using molecular scissors (restriction enzymes, they range to obtain different genes and have different end sequences)

2) Inserting the Gene into a vector (Plasmid)

ainxxx — 2014-04-14 04:38:02 UTC

1) Using DNA ligase to form phosphodiaster bonds with vector so that the DNA fragments would be joined together.

2) Plasmid with DNA insert is called the recombinant plasmid.

3) Putting the recombinant vector into host cells

ainxxx — 2014-04-14 04:45:13 UTC

1) Preparing competent cells for the vector

2) Through different methods:
- Electroporation (cells subjected to high-voltage pulse which temporarily disrupts the membrane and allows vectors to enter cell)
- Bacteriophage (vectors are packaged into phage capsid which will inject vectors into cell)
- Gene gun
- Liposomes

4) Plating of the cells

ainxxx — 2014-04-14 04:49:07 UTC