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15 Cards in this Set
- Front
- Back
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what is recombinant DNA? |
DNA that has been combined |
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how is DNA usually recombined? |
with restriction enzymes that cut DNA at specific sequences sticky ends want to stick because it is complementary form hydrogen bonds ligase fixes nicks |
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what is the purpose of DNA recombination? |
understanding what a gene does making pharmaceuticals making transgenic species sequencing |
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what is a vector and what are its components? |
plasmid, viruses, artificial genes oriC-to get many copies selectable marker-ex. ampicillin-identify cells transformed because bacteria grew -did transformation work restriction sites-put gene inside reporter-tells us if cloning worked (lacZ gene) |
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what are the steps to vector transfer? |
1) get DNA into vector-reporter gene 2) get vector into cell |
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what is Sanger sequencing and how does it work? |
dideoxy NTPs-stops chain elongation has fluorescent tags-each base has a different color electrophoresis using capillaries-smallest appear first |
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what is cDNA and how is it made? |
complementary DNA-complementary to RNA DNA made from mRNA-reverse transcriptase only get mRNAs from genes being expressed-see where genes are being expressed RNA digest with RNAase DNA synthesis with DNA polymerase |
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what is rtPCR used for? |
to quantify DNA |
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how does rtPCR work? |
paired with reverse transcriptase to turn mRNA into DNA PCR amplifies DNA amplification is tracked and quantified using fluorescence |
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what is the end goal of rtPCR? |
collect data used for gene expression exponential |
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how are transgenic animals made? |
added to germ cells-reporter cells |
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what are examples of a transgenic animals? |
glofish-glow in environmental stress -only glow with a promoter livestock that make human proteins in milk |
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what is an advantage of livestock making human proteins in milk and why? |
bacteria is not needed introns are cut out in eukaryotes but not in bacteria |
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how are genes introduced in transgenic plants? |
bacteria, viruses, shooting DNA covered beads, injection into protoplasts (plant cells without cell walls) |
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what do the genes do that are given in transgenic plants? |
resistance to herbicide resistance to insects-BT found naturally in soil improved quality |
