Laboratory Notebook: Cloning Mutant Telomerase RNAs, 1988 - 1989

The notebook begins with a general expression of telomerase RNA through in vitro experiments without extraneous nucleotides.

Also Greider discussed Polymerase Chain Reaction cycles. The cyles are denaturing the protein, spinning down to a nucleate, loading the gel. Greider then began to dye the gel.

Greider was able to draw out the universal primer and reverse primer from the gel. However, found no new mutations in the composite sequence of both.

Greider was also diagramming out the ability of the RNAse enzyme onto certain oligonucleotide sequences. The sequences were then synthesized alongside the G4T2 sequence and then translocated.

Greider conducted a series of southern blot tests and gels. Found that the Library miniprep was degraded. Decided to gather the cells and ran them through a gel, causing them to be diluted. The RNA strand, extracted from plasmids was run through Polymerase Chain Reaction, which amplified the macromolecule that one was using through the enzyme Polymerase, to make mutant substitutes. The process utilizes two reactions. They used two distict 5 prime Mutants in T1 and T2. Greider ran the amplified RNA through a series of gels, however from gel 2-14 the RNA became degraded. Greider found that she still had some DNA left from prep that she made from sequencing. The notebook conlcuded with two items. The first are a list of restriction enzymes on certain DNA sequence, corresponding diagrams on plasmid DNA (106 kb). The second is a publication by Brow and Guthrie (Yeast U6 snRNA is remarkably conserved) and its corresponding figures.