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Fantastic Summary of Methods in Molecular Biology for Entire Course.pdf

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BIOL 309
Barbara Moffatt

Technique Research use Controls Benefits Limitations Genetic approaches To identify Must first Can identify Best done on Forward genetics mutants affected define the genes involved model in a specific normal range in a process organisms process or of response in that you never that are small response the wild type would have in size small population expected genomes low control is excellent repetitive testing many starting point DNA cheap genetically to understand to maintain identical the molecular diploid individuals basis of a and selecting a process or mutant response outside of this range Methods to Make multiple You can learn Must be able Reverse genetics reduce the transformants the phenotype to transform gene silencing expression of of the of an organism the organism specific genes or construct and that lacks of interest gene families if show that they expression of a need to be the sequences of are similar target gene of thoughtful in these genes are show that the interest without the design of known expression of having a true the silencing the target mutant can be construct so gene is fast can reduce it doesnt reduced as expression in target nonexpected all cells or a specifically few cells of the organism depending on the promoter used Gene cDNA recovery techniques Many uses Show the This is way to Have to have Restriction including identify uncut sample orient the enzyme mapping specific DNAs is a different sequences on a needed and determine size if doing large piece of be able to direction of double DNA you can determine the inserts in a vector digests do the describe the sizes of the singles too location of a fragments sometimes fragment well using verify enzyme relative to appropriate works by specific size markers cutting a restriction sites fragment you know has the siteTo separate Must evaluate Useful for Some Genomic library fragments of a the quality of screening with fragments are genome into the library by heterologous not wellsegments that can estimating the probes stable represented be maintained average insert source of in the library easily and shared size and the genomic particularly with others total number fragments repetitive cloned in phage of used for DNAs very or plasmid vectors recombinants genome large in the library sequencing and genomes tells you if all mapping require many the fragments clones in the genome are in your collection To separate Evaluate the Used for a large Reverse cDNA library copies of mRNAs number of number of transcriptase into individual recombinants applications is not highly clones that can be in the library depending on processive so stored longterm to estimate type of vector some clones and shared how well you expression will be have captured vectors used partial hard copies of all for to clone very the mRNAs in complementatilarge very the sample on small or very overexpression rare cDNAs EST collections virtual northerns cDNA is the means for storing and analyzing mRNA which
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