Term
| what are restriction enzymes more accurately known as? |
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Definition
restriction endonucleases
Restriction enzymes are more accurately described as restriction endonucleases as they act as nucleases (enzymes that attack nucleic acids) within (endo) the nucleic acid structure. This is in comparison to an exo-acting enzyme that attacks a structure from the end of the molecule. |
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Term
| what is commonly used as a vector? |
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Definition
a plasmid
A plasmid is an extrachromosomal piece of DNA that can be easily isolated from bacteria to be used as a vector |
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Term
| what is the most common way to select bacterial cells that have taken up a plasmid? |
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Definition
antibiotic resistance
Antibiotic resistance is encoded by a gene carried on the plasmid. Hence only those bacteria transformed with the plasmid will be able to grow on agar plates containing the appropriate antibiotic. |
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Term
true or false
most genetic engineering experiments involve restriction enzyme digestion of the entire bacterial chromosome |
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Definition
false
Bacterial chromosomal DNA is not normally used in genetic engineering experiments |
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Term
| which technique involves amplification of DNA? |
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Definition
PCR
The polymerase chain reaction amplifies a piece of DNA using sequence specific primers |
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Term
denaturing DNA fragment to be amplified
annealing of primers complimentary to the sequences
primer extension with DNA polymerase
Repeating the 1st 3 steps in many cycles are all steps in which reaction?
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Definition
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Term
| what is needed to carry out DNA sequencing reaction? (5) |
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Definition
| ssDNA, dNTPs, ddNTPs, primer and DNA polymerase |
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Term
| What us the relevance of Northerm blotting? |
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Definition
| To gain information about when a gene is transcribed and the size of a transcript but not the number of genes encoding a particular protein |
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Term
| the identification a protein within a complex mixture is carried out using western analysis. this includes the use of?... |
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Definition
SDS polyacrylamide gel electophoresis
blotting
2 antibodies |
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Term
| synthesis of cDNA from mRNA using PCR requires the combination of which enzymes? |
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Definition
| reverse transcriptase and thermostable DNA polymerase, |
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