21.1 producing DNA fragments Flashcards Preview

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Flashcards in 21.1 producing DNA fragments Deck (14):
1

what are the 5 stages of making a protein using the DNA technology?

1. isolation
2. insertion
3. transformation
4. identification
5. growth/ cloning

2

what is recombinant DNA?

DNA of 2 different organisms that have been combined

3

what are transgenic organisms?

organisms that have recombinant DNA

4

what are genetically modified organisms?

organisms that have recombinant DNA

5

what are the 3 methods of producing DNA fragments?

1. mRNA -> cDNA using reverse transcriptase
2. using restriction endonucleases to cut fragments
3. creating the gene using a gene machine

6

where does reverse transcriptase come from?

retroviruses e.g. HIV

7

what does reverse transcriptase catalyse?

mRNA to cDNA

8

what is the method for using reverse transcriptase to get a DNA fragment?

1. obtain mRNA from a cell that readily produces the protein
2. add reverse transcriptase
3. cDNA is made
4. add DNA polymerase
5. DNA made

9

what does restriction endonuclease do?

cuts DNA in straight or staggered ends

10

what are staggered ends also called?

sticky ends

11

why is a restriction endonuclease that recognises a 50 base pairs better than a restriction endonuclease that recognises 6?

- more specific
- as statistically appears less often

12

what are straight ends also called?

blunt ends

13

where does restriction endonuclease come from?

bacteria

14

what is the method that involves a gene machine?

1. feed desired nucleotide sequence into the computer
2. nucleotide added one at a time to form a oligonucleotide
3. oligonucleotides overlap and are joined together to make a gene
4. made double- stranded using PCR
5. replicated DNA forms using PCR
6. gene inserted into plasmid as has sticky ends