Module 7 - Microbial Genetics Flashcards
What term describes the determination and study of complete genomic sequences?
Genomics
The entire genome of an organism is called?
Genomic library
In what way have researches relied on genomic libraries for genetic analyses?
Determining sequences of the cloned fragments
The basic unit of heredity that occupies a specific location on a chromosome is called?
Gene
The entire set of genetic instructions found in a cell is called?
Genome
Which sequencing takes advantage of the primer-specific DNA synthesis activity of DNA polymerase?
Sanger or dideoxy sequencing
What are the three steps in Sanger sequencing?
- Cloning of the fragment to be sequences
- DNA synthesis
- Gel electrophoresis
How does Sanger sequencing differ from PCR?
For Sanger sequencing, only a single primer is used in the reaction compared to two
What purpose does Sanger sequencing serve in?
Research purposes, such as targeting smaller genomic regions in a larger number of samples
- Low error rate
Which sequencing method is based on the synthesis of a DNA strand, using a template strand to guide the insertions of nucleotides
Sanger Sequencing
Which method of sequencing involves first sequencing the cloned DNA as far as possible then, the newly obtained sequence information is used to design another sequencing primer
Primer walking
Which method of sequencing is similar to Sanger sequencing in that it relies on DNA polymerase-mediated synthesis of the complementary strand of a single-stranded template?
Pyrosequencing
When is a dNTP incorporated into a growing piece of DNA in pyrosequencing?
Only if the dNTP is complementary to the template base
What is released when the dNTP is incorporated into DNA, in pyrosequencing?
Pyrophosphate (PPi)
Which enzyme converts pyrophosphate into ATP in pyrosequencing?
ATP-sulfurylase
After ATP-sulfurylase takes place, what occurs (pyrosequencing)?
Production of light
Which method of sequencing shears DNA into short pieces and then sequences those fragments from their ends
Shotgun sequencing
Which method of sequencing is also referred to as next-generation sequencing?
High-throughput sequencing
Which method of sequencing breaks DNA into short pieces, amplified, and then sequenced?
High-throughput sequencing
Which method of sequencing does not break down or amplify DNA but instead directly sequence a single DNA molecule?
- DNA read length
Third-generation sequencing
Advantages of primer walking?
Quicker and easier method of sequencing long stretches of DNA
Examples of high-throughput sequencing?
- Pyrosequencing
- Illumina sequencing
- PacBio
When is shotgun sequencing useful?
Used to remove errors, fill gaps or correct parts of the sequence that were originally assembled incorrectly
What is the study of the entire set of genes in the genome of a cell
Genomics
What is the study of the entire set of proteins produced by the cell
Proteosomics
What is the study of the entire set of mRNA synthesized by a particular organism
Transcriptomics
What is a two-dimensional separation method that separates based on BOTH isoelectric point and mass
2D-PAGE
What properties of proteins does 2D-PAGE utilize?
As they migrate along the gradient, they reach a point on the strip where there is NO net charge
- pH equals pI of the protein
What identifies the unique mass-to-charge ratio of proteins that helps reveal their identity by comparison of similar ratios identified for known proteins?
Mass spectrometry
Two methods used to determine protein structure?
- X-ray crystallography
- Nuclear magnetic resonance
