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Chapter 20
DNA Tools and Biotechnology
| Question | Answer |
|---|---|
| biotechnology | the manipulation of organisms or their components to make useful products |
| nucleic acid hybridization | the base pairing of one strand of a nucleic acid to a complementary sequence on a strand from a different nucleic acid molecule |
| genetic engineering | the direct manipulation of genes for practical purposes |
| DNA Sequencing | the DNA first cut into fragments, and then each fragment is sequenced |
| Next-generation sequencing (Steps 1-4) | - DNA is fragmented -Fragments are isolated from aqueous solution -PCR copy, using template strand -bead placed into small well with DNA Polymerase & primers to hybridize 3' strand |
| Next-generation sequencing (Steps 5-8) | - Different sequenced DNA fragments -nucleotide joins growing strand -nucleotide is washed off and a different nucleotide is added - washes are repeated; reveals sequence;completed complementary strand |
| DNA cloning | preparing well-defined segments of DNA in multiple identical copies |
| plasmids | small,circular DNA molecules that are replicated |
| recombinant DNA | a molecule containing DNA from two different sources, very often different species |
| gene cloning | the production of multiple copies of a single gene is a type of DNA cloning |
| cloning vector | DNA molecule that can carry foreign DNA into a host cell and be replicated there. |
| What are two purposes of gene cloning? | to make many copies of, or amplify a particular gene and to produce a protein product from it. |
| What are the four steps to Gene cloning? | - Gene inserted into plasmid (vector) -Plasmid in bacterial cell - Host cell cultures clone of cells with gene of interest - applications |
| what do restriction enzymes protect? | bacterial cell by cutting up foreign DNA from other organisms or phages |
| Using restriction enzyme and DNA ligase to make recombinant DNA plasmid | -restriction enzyme cuts sugar phosphate backbones at each arrow -DNA fragment from other sources are added. Base pairing of sticky ends produces various combinations - DNA ligase seals the strands |
| The key to the high specificity is the pair of __ used for each PCR amplification | primers |
| stem cell | relatively unspecialized cell that can both reproduce itself indefinitely and, under appropriate conditions, differentiate into specialized cells of one or more types |
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