1. Recombinant DNA Daredevils Yen Phan, Jen Masciovecchio, Cristina Johnson, Praj Acharya, Julie David

2. Cloning  Cloning is used by scientists to study a particular gene that they are interested in  Bacteria have vectors that are used to store DNA information such as antibiotic resistance gene.  The origin of replication within a vectors signal DNA replication, so we can use it to clone genes we want.

3. Cut &Paste  In addition, through evolution bacteria created different types of restriction enzymes used to catalyze reaction. It catalyzes the breaking of phosphodiester bond in a specific sequence.  So basically, Each restriction enzyme identify the nucleotides of interest and cut it at specific sites.

4. Vector &Transformation  When mixed together, both the vector and the DNA strand of interest, would joined through a hydrogen bond. The ligase used to bond the okazaki fragment in lagging strands of DNA is then used to covalently tape the pieces together.  To defend itself from cutting it's own DNA, the bacteria has a methylation group in the site where the restriction enzyme can't cut it.  When we mixed the vector with the bacteria, the bacteria would slurp the vector in. This is called horizontal transfer when bacteria shared DNA with each other to make sure that the genes best fit for the environment get passed on. If we stock them with heat and cold. It will take the vector in at a faster rate.

5. Selection  Then, the bacteria is placed in a petri dish with nourishing food for them to reproduce. All the bacteria with or without the vector will clone and create a colony around it.  This is known as the library where it has millions of different bacteria with different vectors and different genes stored in it.  Since the bacteria with the vector already have the antibiotic resistance gene, we just have to put ampicillin, penicillin, or other antibacterial drug on the dish and it will kill most of the bacteria. The ones that take in the selected vector will live.  Next, the bacteria is placed in a plasmid prep to break up the vector from the bacteria.

6. Gel Electrophoresis  The vector is placed in an agarose or a jello bar. On one end is a negative charge while the other is a positive charge. Since DNA is negatively charged, it will go to the positive end. Smaller DNA will move further.  Also, there will be a molecular weight ladder used as a ruler to mark where the base cut would be in the vector.  The band is where you see the line or where the cut would be. You can use the band and ruler to see how many cuts take place and where each cut is or how close they are.

7. Questions  How does some viruses obtain their DNA methylation group in the first place? The explanation was not very clear in the video.  How do we know what restriction enzyme to use and where it cut in the gene in the first place?  Is the shorting of the gel electrophoresis based on the charges or the weight?  How do they know which antibiotic restriction gene to put into the vector?