RECOMBINANT DNA By the White Coat Wonders Megi Resulaj & Dalena Dang

TOPIC STATEMENT #1 Bacterial plasmids are used to carry and transfer a specific gene to an organism. They must contain an origin of replication in order for the plasmid to be integrated and replicated inside the host cell.

TOPIC STATEMENT #2 Restriction Enzymes cut the backbone of DNA at a specific sequence and are used to manipulate DNA (example: lab work with pGLO).

TOPIC STATEMENT #3 Certain restriction enzyme sites may be present in a certain allele (either the affected or unaffected allele). When an organism is heterozygote for a disease, i.e. Aa, under PCR the DNA will split into the designated lengths where the restriction site is present. In addition it will include the original length without the split from the allele that is not affected by the restriction site. As for homozygotes, i.e. AA or aa, each would respectively express either only the split DNA fragment or the full desired DNA fragment.

TOPIC STATEMENT #4 A Polymerase Chain Reaction replicates and amplifies fragments of DNA, usually a specific gene, using primers and dNTPs.

TOPIC STATEMENT #5 Electropheresis is a method of estimating base pair lengths of certain fragments in DNA, which travel through agarose, an intricate mesh-like structure. This mesh like structure allows for shorter fragments to travel further down the gel, and the longer pieces stop traveling closer to the top of the gel.

QUESTION #1 In the handout, there were several restriction enzymes that cut at the same sequence. Is it possible to know/tell if one restriction enzyme is more favorable than another?

QUESTION #2 What are the bands in the gel electrophoresis? Do the DNA fragments travel vertically? Why are all the band the same size?

FOLLOW-UP Treating Genetic Diseases Can we combine genes from other species (even though they might species specific) with the human genome, which a human would then be able to express and gain a specific trait that could better human function?