Term
| What do restriction enzymes naturally do in prokaryotes? |
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Definition
| Protect them from hostile foreign DNA |
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Term
| ____ restriction enzymes cleave DNA within their recognition site |
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Definition
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Term
| Six steps of molecular cloning |
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Definition
1) Isolation and fragmentation of source DNA with restriction enzymes
2) Insertion of DNA fragments into a cloning vector
3) Incorporation of selected DNA fragment into host cell
4) Screening of host cells for clonal insert
5) Detection and purification of desired clonal cell line
6) Production of large number of cells with desired clone to make product |
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Term
| Maximum size of inserted DNA in bacteriophage cloning vector? |
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Definition
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Term
| Name 4 ways genetic engineering differs from natural genetic/evolutionary mechanisms. |
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Definition
1) Increases size of genomes with genes that normally would never develop naturally in some species
2) Genes cross species boundaries without loss of reproductive viability
3) Speeds appearance of new characters in species compared to natural mutations
4) Man-made selection process involved instead of natural selection process |
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Term
| The 454 pyrosequencing method is ___ times faster than Sanger sequencing |
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Definition
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Term
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Definition
DNA broken into small ssDNA (100 bases each)
Each ssDNA immobilized to a bead and put in a separate plate well
DNA amplified to make several copies of DNA on each bead
Nucleotides flooded one by one over plate wells
Each time a nucleotide is incorporated into the complementary DNA strand pyrophoshate is released which provides energy for a light pulse
Each light pulse signals the incorporation of a base complimentary to one in original DNA at that point and DNA sequence in revealed
Sequences of small DNA sequences assembled into entire genome sequence using sequence overlaps as guides |
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Term
| Routine size of oligonucleotides for DNA synthesis? |
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Definition
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Term
| Steps in de novo synthesis of small DNA fragments? |
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Definition
Desired sequence submitted
First nucleotide in sequence fastened to porous glass beads
After each elongation step, reaction mixture flushed out of beads
Next steps proceed with new reaction mixture and appropriate nucleotide |
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