Answers | Verified | Latest 2024 Version
What is genetic engineering? - ✔✔using in vitro techniques to alter genetic material in the lab
What is recombinant DNA technology? - ✔✔the artificial recombination of DNA from two organisms
What are some basic techniques of genetic engineering? - ✔✔
DNA amplification
electrophoesis
nucleic acid hybridization
molecular cloning
expressing foreign genes
targeted mutagenesis
What is heterologous expression? - ✔✔expressing a gene in a different host
What is the polymerase chain reaction? - ✔✔rapid amplification in the number of copies of specific DNA
sequences for further analysis
What is a thermocycler? - ✔✔an automated PCR machine
What is quantitative PCR? - ✔✔quantifies initial amount of DNA
What are some applications of PCR? - ✔✔-obtain specific fragments of DNA for cloning
-comparative (phylogenic) studies to amplify genes and DNA sequences
-amplify VERY small amounts of DNA (forensic)
,What are the steps in PCR amplification? - ✔✔1. DNA is denatured by heating
2. Synthetic piece of DNA flanking sequence of interest is added
3. Add DNA polymerase (Taq or Pfu polymerase)
4. heat and cool - repeat
Why is use of thermostable DNA polymerase critical? - ✔✔because of high temperatures
What do PCR require? - ✔✔DNA polymerase and artificial oligonucleotide primers made of DNA
What is reverse transcription PCR? - ✔✔can make DNA from an mRNA template
What enzyme is used in reverse transcription PCR? - ✔✔reverse transcriptase
- converts RNA into complementary DNA
What is gel electrophoresis? - ✔✔the separation of charges molecules using an electric field
What are agarose gels stained with? - ✔✔ethidium bromide
DNA cut with different restriction enzymes will have the same or different banding patterns on an
agarose gel? - ✔✔different
What is nucleic acid hybridization? - ✔✔Single strands of DNA, or of DNA and RNA, from related
organisms will hydrogen bond to form a double stranded molecule
What are nucleic acid probes? - ✔✔ss DNA fragment complementary to the gene of sequence of interest
How are nucleic acid probes created? - ✔✔cloning, synthesis, denaturing a fragment of DNA
, What is a southern blot? - ✔✔a hybridization procedure where DNA is in the gel and probe is RNA or
DNA
What is a northern blot? - ✔✔RNA Is in the gel and the probe is DNA or RNA
What is FISH? - ✔✔Fluorescence in situ hybridization (FISH) uses fluorescent probes attached to
oligonucleotide
What is molecular cloning? - ✔✔movement of a gene from original source to small genetic element
(vector) which can be manipulated
What are the main steps of gene cloning? - ✔✔1. isolation and fragmentation of source DNA
2. Insertion of DNA fragment into cloning vector
3. Introduction of cloned DNA into host organism
What are restriction endonuclease enzymes? - ✔✔tool in genetic engineering that recognizes specific
DNA sequences and cut DNA
What do type II restriction enzyme endonucleases do? - ✔✔-cleaves DNA within recognition sequence
-most useful for specific DNA manipulation
What do type I restriction enzyme endonucleases do? - ✔✔-first discovered + purified
-cuts DNA at random for from recognition sequence
What do type III restriction enzyme endonuclease do? - ✔✔-large, combination restriction and
modification enzymes
- cleaves outside of recognition sequences
Structure of cleaved product from type II REases may differ how? - ✔✔3' or 5' overhang and blunt ends