Week 1: Fragmenting DNA w/ RE
Sequences that are read the same forward and backward
Transformation
This is a lab technique that amplifies small segments of nucleic acids
PCR
Biology technique that separates DNA based on size
Gel Electrophoresis
These are the differences between + and - controls
+ control: used to show a positive test result
- control: used to show a negative test result
These cleave (cut) the backbone of DNA molecules at palindromes
Restriction Enzymes
Plasmids
Used in PCR to break open cells to release their components for DNA isolation
Lysis Buffer
Labels used for a size standard for comparing DNA bands
The variety of different viruses that can attack the same cell type refers to
Viral cell specificity
A specific place on DNA molecule where a restriction enzyme cuts
Restriction enzyme sites or Restriction sites
This gene was transformed into the E. coli MM294
ampr
This breaks down proteins that could interfere with amplification, including those around the DNA molecule and within the cell
Proteinase K
These are formed on gels by many pieces of DNA that are the same and differing sizes
Bands or Restriction Fragments
These are the differences between selective and differential media
Selective: Used to selective for the growth of a particular microorganism, inhibiting the growth of others
Differential: used to differentiate closely related organisms, due to presence of dyes/chemicals in media
Where the larger molecules are found in an agarose gel
Towards the top
Penicillin (Ampicillin in our lab) kills bacteria by blocking the synthesis of
Peptidoglycan in the cell wall during cell division
These were found inside of the PCR pellet
Taq DNA polymerase, dNTPs, MgCl2, and buffer
TBE (electrophoresis) buffer is used for these two reasons
Provide ions that carry the current to move DNA bands, separate nucleic acids, and provide a stable pH environment
This is used to describe the number of different strains or species of organism that a specific virus can infect
Host cell specificity
The type/name of cut restriction enzymes EcoRI and HindIII leave on DNA
Sticky ends
The steps that are considered critical to the process of transformation
Initial ice incubation, heat shock, secondary ice incubation, and room temperature incubation
AKA Preincubation, Incubation, Heat shock, and Recovery
These are the three steps of PCR and what they do
Denaturation: Heat DNA to a high temp that breaks the hydrogen bonds between base pairs
Annealing: Temp is lowered to allow primers to bind to the template strands
Extension: DNA polymerase adds NTs to the 3' end of the primer
Loading dye is used in gel electrophoresis for these two reasons
Allow for color DNA visualization and increasing the density of DNA
The difference between Gram + and Gram - bacterium
Gram +: stain purple, thick layer of PTG in cell wall
Gram -: stain red/pink, thin layer of PTG in cell wall