DNA Manipulation Flashcards
DNA manipulation
using DNA in a lab for our benefit
Gene editing
Process of making targeted changes to a specific section of an organism’s genome, including adding, deleting or altering specific DNA sequence
Restriction Enzymes (endonucleases)
Enzymes that cut DNA at a specific location (restriction site)
Palindrom
a sequence of DNA, that you can read from 5’ to 3’ and vice versa
Polymerase
joins base pairs together
Helicase
Unzips DNA
Gyrase
Unwinds DNA
RNA polymerase
makes a polymer of RNA
DNA polymerase
makes a polymer of DNA
Reverse transcriptase
makes cDNA from mRNA
endonuclease
cuts DNA at a specific recognition sequence
Ligase
Joins DNA segments with “sticky ends”
Gene probes
Single-stranded DNA or RNA complementary base sequences to target DNA. Attached with fluorescent markers. DNA must denature and then anneal
CRISPR
cas9 are enzymes in the cell that can cut up DNA that matches a little piece of DNA that they hold inside of themselves.
Cas9 will take the RNA transcribed from the DNA and hold the RNA in the enzyme to travel around the cell to find DNA that is complementary to the RNA.
This can be used to cut DNA at any place we want which lets us alter the genome
Ligase
Opposite of endonucleases. Enzyme that can stick two single strands of DNA together to form one longer strand
DNA amplification
Using the technique called polymerase chain reaction (PCR), researchers can create vast quantities of DNA identical to trace samples
Polymerase Chain Reaction
Technique by which one can copy segments of a DNA sample repeatedly to form multiple copies of the same DNA for further analysis
PCR process
- mixture of DNA fragments heated to seperate strands (92 degrees)
- temperature cool to allow primer to bind to DNA
- temperature increased again to allow Taq DNA polymerase to start building complementary strand
- process is repeated over and over
Gel Electrophoresis
method of separating fragments of DNA, usually according to size
Gel Electrophoresis process
- sheet of gel used to seperate fragments
- DNA fragments added to one end of gel
- electric current run through gel
- electric current run through gel
- DNA is negatively charged
Plasmid
double-stranded circular DNA molecules separate from the bacterial chromosome. Replicate independently. Usually has its own marker gene
Recombinant plasmid
plasmids that we put foreign genes into. Some plasmids may not incorporate the gene of interest because they may reseal before the gene of interest can bind. This is because the complementary sticky ends are attracted to themselves
Transformed bacteria
bacteria that have incorporated the recombinant plasmid
Electroporation
using electrical currents to open pores and make the membrane permeable
