Lecture 17 Flashcards
(25 cards)
Restriction Endonuclease
Cuts DNA at a specific sequence
Palindromic Site
When reading from the 5’ to 3’ end the complementary strand reads the same as the original strand. Ex: 5’ GAATTC3’ : 3’CTTAAG5’
DNA Ligase
Puts the cut ends back together. Remakes phosphodiester bonds
Complementary ends
The other strand of DNA that is paired to the parent strand
Recombinant DNA Molecule
DNA molecules formed by laboratory methods of genetic recombination to bring together genetic material from multiple sources, creating sequences that would not otherwise be found in biological organisms. Uses DNA ligase
Electrophoresis
Separates DNA fragments based on size. It is a gel and large fragments stay at the top and small fragments go to the bottom
Labeled Probe
Marks the DNA so it knows where the probe is. It can be radioactive, emit light, or be a color dye. Probe is often used as a replication primer. The longer the probe the more specific DNA it binds to
Hybridization
Measures how similar the DNA are to each other
Mutagenic Primer
One of the bases are wrong and it makes the sequence mispair. A few mismatches are okay.
DNA Library
Restriction enzyme(cuts DNA) can be used to generate a DNA Library. a collection of DNA fragments that is stored and propagated in a population of micro-organisms through the process of molecular cloning
Genomic Fragments
When the DNA is cut into pieces the pieces are called genomic fragments.
Cloning Vector
a small piece of DNA, taken from a virus, a plasmid, or the cell of a higher organism, that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes
Host Cell
The cell that holds the DNA from the cloning vector
Colony Hybridization/ Library Screening
It starts in a plate and you put a screen over it and the colony is transferred to the screen and you add nucleic acid and then filter it and the colony is developed and the specific colony you are looking for stands out
Reporter Gene
Reports if a gene is turned on or off. Allows easy detection of transcription from a promoter
Green Fluorescent Protein (GFP)
If it is on it turns green and if it is off there isn’t a color.
Southern Blot
Uses probes to find DNA fragments in an electrophoretic gel
Polymerase Chain Reaction (PCR)
Uses special primers to amplify a specific piece of DNA. Denatures the DNA, Binds specific primers to the DNA, Makes new DNA from the primers.
PCR Screening (for diseases)
Use PCR to check to see what is the DNA of a person. Can see if something is wrong with them
Gene Cloning
It is like making a library, except only a specific gene is inserted into the new host cell
Transgenic/ Genetically-modified organisms
An organism whose genetic material has been altered using genetic engineering techniques
Pyrosequencing
The sequence of any DNA or RNA can be rapidly determined, even while replicating
Sequence “reads”
Uses light and the bases of DNA to see what comes next in the sequence. PPi is released and AMP is added and ATP is made and then luciferase is added to create light and then if it isn’t there the base is washed away with apyrase and the next base is added.
Genome Assembly
Puts a whole DNA sequence together. It matches up the small fragments and finds where they overlap. It also takes snapshots to each piece and they are laid on top of each other.
