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what does topoisomerase do? - ANSWERbreaks phosphate backbone to "un" supercoil DNA Another name for Topoisomerase II - ANSWERGyrase Helicase - ANSWERUn"zips" DNA at replication fork What makes up the primosome? - ANSWER1. primase 2. helicase 3. DNA C NASBA stands for... - ANSWERnucleic acid sequence based amplification NASBA - ANSWERRNA is target RNA--cDNA Reverse transcriptase (RNA--cDNA) followed by DNA dependent polymerase (cDNA-- RNA) RNAse H degrades original RNA template Evaluated gene EXPRESSION - ANSWERRNA ratio for organic extraction - ANSWERPhenol (50): Chloroform (49): isoamyl alcohol (1) phenol - ANSWERdenatures proteins, dissolves lipids chloroform - ANSWERassists in dissolving remaining lipids, gives a clear interphase between aqueous phase Long term storage of RNA - ANSWER70% EtOH at -70 C ratio for 28S to 18S subunits - ANSWER2:1 (can see this in a gel) A260 - ANSWERnucleic acids A280 - ANSWERprotein A230 - ANSWERorganic compounds, phenol A270 - ANSWERphenol optical density of ds DNA - ANSWER50 ug/ml optical density ssDNA - ANSWER33 ug/ml optical density RNA - ANSWER40 ug/ml DNA concentration is calculated: - ANSWERA260 x dilution factor x 50 ug/ml RNA concentration = - ANSWERA260 x dilution factor x 40 ug/ml DNA B - ANSWERhelicase DNA G - ANSWERprimase Processivity - ANSWER# of dNTPs that a polymerase can incorporate continuously without dissociating from primer template fidelity - ANSWERerror rate of polymerase star activity - ANSWERRE : additional cleavages at imperfect recognition sites synthesis of DNA is in what direction: - ANSWER5'--3' add to 3' -OH Taq DNA polymerase - ANSWERthermus aquaticus thermostable used in amplification 3 termination sequences - ANSWERUGA UAA UAG Huntingtons Disease (genetic basis) - ANSWERAutosomal Dominant Chr 4p16 IT-IS gene gene (Huntington Protein) CAG repeats 29 = normal 29-35= borderline 35 = Huntington Disease 100% penetrance Huntington's disease phenotype - ANSWERdegeneration of brain cells uncontrolled movements loss of intellectual faculties emotional disturbance Cystic Fibrosis - ANSWERAutosomal recessive CFTR gene most common inherited form of mental retardation - ANSWERfragile X Fragile X - ANSWERFMR 1 gene on Xq27.3 CGG repeat normal = 5-44 carriers= 59-200 fragile X- over 200 Hereditary hemochromatosis - ANSWERiron overload disease autosome recessive HFE gene SARs stands for - ANSWERsevere acute respiratory syndrome 6 open reading frame for Sars-cov19 - ANSWER1. replicase enzyme 2. spike protein 3. envelope 4. membrane 5. glycoproteins 6. nucleocapsid protein Ct = - ANSWERthe point at which fluorescence crosses the threshold Nick translocation - ANSWERmethod of probe labeling -ss nicks in DNA -DNApol I adds nts to the nick -1 nt is labelled random labelled primers - ANSWERlabeling probes