Lab 9 and other questions Flashcards
bioinformatics
a combination of molecular biology and computer science that aims to develop the technology for organizing, analyzing, and storing sequence data
Difference between accuracy and precision?
Accuracy- close to true value
Precise- close to each other
What are you digesting?
Sample DNA
Do you understand plasmid and linear DNA restriction/ligation? See Extra Help, Lab 6 folder
What are we ligating? What products could form during ligation
Sample DNA and plasmid. Recombinant/nonrecombinant plasmid
What are the steps of transformation? (What happens at 42ºC?
heat shock
Can also have E.coli with nonrecombinant plasmid or no plasmid… how do we distinguish?
How and why do we purify our products? Hint: compare PCR and Sequencing Reactions
Purpose of Alkaline Phosphatase?
Remove phosphate groups, ______________-
What are the sequencing reagents? Purpose of each reagent?
Purpose of Blastn?
Nucleotide search and alignment program
Blastx?
Protein database
Blat?
DNA search to find homologs between closely related species (fast, but less accurate than blastn/x)
annotate the gene sequence
RNA and amino acid sequences it encodes are determined
comparative genomics,
information such as the sequence, function, and location of a gene are evaluated across different species
homologous.
genes that are evolutionarily related to one another
How does Sanger Sequencing work?
- template DNA is targeted using a sequencing primer.
- Primer binds to DNA adjacent to region of interest.
- Chemical analogs of the four nucleotides in DNA are used.
- DNA replication occurs, and the sequence of nucleotides is determined.
Why add a really rich broth?
What happens at 37ºC?
Bacteria recover, replicate the new DNA (plasmid), and express the desired gene
How do you calculate TE? What does TE tell us?
