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| small DNa molecule that can replicate independantly of chromosomal DNA |
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| can be present in many copies in a cell |
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| can only be present in a couple of copies in a cell |
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| has resistance so growth selects for clones cells |
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| contains origins of replication for multiple cells |
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| why don't restrction enzymes cut a bacteria's DNA |
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| has been cut and linked back together without incorporation of new DNA |
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| non metabolizeable lactose, those with functional gene will cleave X-gal and appear blue due, those without functional gene will not cleave XGal and will remain white |
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mRNA
add poly A primer for 3' tail,
add reverse transcriptase and bases to make DNA
RNase H digests some of the RNA, leaving RNA primers on the other side so DNA polymerase/ligase can make and seal the other side of the DNA |
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| shorter, much easter to sequence, has no introns, only genes of interest |
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| thermally stable form of DNA polymerase |
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| denature DNA, anneal primers to DNA, primer extensions with Taq polymerase repeat |
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| regulate temp and timing of PCD, typically 20 to 30 cycles |
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| reverse transcriptace PCR |
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| first round is RNA to DNA |
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| compares DNA in electrophoresis gel, uses ethidium bromide to stain DNA, and UV light to compare size |
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| has that weird TaqMan detector that emits a fluoresence but has a silencer, anneals to a DNA sequence, when the DNA is synthesized it kicks the detectr off and it goes away from the silencer so it emits a fluoresence, fluoresence is measured against the background level to find the original amount of DNA |
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| collection of vectors when the original statring material was a chromosome |
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| recombintnat library made of cDNA pieces that were transcribed from mRNA, contains only genes, no introns |
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| master plate, use detergants and NaOH, use fluorescently labeled DNA, see which bacteria colonies it anneals to look under XRay to see which bacteria have gene you want |
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| can detect particular sequences in a DNA genome |
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| Used to detect a specific RNA in a pool of RNA molecules; can tell you the size and relative quantity of a particular RNA molecule in cells from different tissue types |
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| Used to detect proteins; involves antibodies as probes to bind to a particular antigen on a protein of interest |
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| Used to screen DNA libraries for a gene of interest; involves a nylon membrane and radio-labeled probes |
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| studies DNA protein interactions, DNA with bound protein will not travel as far as DNA without, figures out where the protein binds |
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| no 3' OH end on dNTPs so more can't be added is stopped |
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| site directed mutagenesis |
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| base is mutated, may or may not be reparied, if it is then new different proteins will be made |
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| DNA from another organism has been introduced into an organism |
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