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Add to wolbachia notes!MONDAY IS A BIG LAB! BE READY!
DNA EXTRACTION REAGENTS
1. Phosphate Buffered Saline (PBS): a salty solution of constant pH
to keep tissues, cells, and proteins intact during maceration
2. Proteinase K: an enzyme that catalyzes the breakdown of
cellular proteins by splitting them into smaller peptides and amino
acids
3. Buffer AL: a cell lysis solution that breaks open cell and nuclear
membranes
4. Ethanol: used to precipitate DNA from the extracted material
5. Buffer AW1 and AW2:
solutions that wash the DNA attached
in the column membrane of contaminants
6. Buffer AE: a solution that elutes the DNA from the membrane
and allows stable storage of DNA for years in the refrigerator or
freezer
MONDAY IS A BIG LAB!!!! BE READY!!!
WOLBACHIA PCR
We are amplifying part of the 16S ribosomal RNA gene
that is unique to wolbachia. It is 438 bp.
We are also amplifying a portion of the gene for
cytochrome c oxidase. It is 650 bp. This is a gene
specific to animals, not bacteria. Why would we do this?