MB (ASCP) Exam Test Questions And Answers Verified 100% Correct

MB (ASCP) Exam Test Questions And Answers Verified 100% Correct 3 steps of PCR - ANSWER1. denature 2. Anneal 3. extension temperature for denature stage of PCR - ANSWER90-95 C temp for annealing step PCR - ANSWER40-60 C temp for extension phase PCR - ANSWER70-75 C what makes a good primer? - ANSWER18-28 nts in length 50-60% GC content Tm of primer sets are within 5 C of each other Tm= - ANSWER4(G+C) + 2(A+T) ligase chain reaction (LCR) - ANSWERrepeated cycles of probe/primer hybridization and ligation to make multiple copies of the target target sequence is NOT duplicated but provides a template for probe hybridization after probe hybridizes, ligase joins the probes terminal denaturation makes the original template and probe available in the next round ligation of primers only occurs if NO MUTATION present TMA stands for - ANSWERtranscription mediated amplification TMA method - ANSWERproduct= RNA uses 2 primers: 1. RNA po l 2. region complementary for target sequence Uses 2 enzymes; 1. Reverse transcriptase to extend primer making cDNA 2. RNA polymerase - makes RNA copies of DNA t(9;22) - ANSWERBCR-AbL t(15;17) - ANSWERPML-RARA t(8;21) - ANSWERRUNX1-RUNX1T1 inv(16) - ANSWERCBFB-MYH11 t(16;16) - ANSWERAML-ETO define heteroduplex - ANSWERa double stranded DNA formed by annealing two ss strands together NASBA - ANSWERNucleic Acid Sequence Based Amplification -isothermal -targets RNA -seq. specific primer with T7 pol promoter. degrades template RNA with Rnase H. Extension by Reverse Transcriptase 2 methods of detection for qPCR - ANSWER1. SYBR Green 2. Modified oligonucleotide probes which fluoresce when hybridize with complementary DNA (quencher, reporter) SYBR Green - ANSWERfluorescent dye which intercalates with dsDNA fluorescence increases as product accumulates with each amplification what is electrophoresis? - ANSWERthe separation of proteins or nucleic acids on the basis of size 3.5% polyacrylamaode gel - ANSWER100-1000 bp 5% polyacrylamide gel - ANSWER75-500 bp which type of gel has higher resolution? - ANSWERpoyacrylamide which type of gel has a greater range of separation - ANSWERagarose size range for agarose gels - ANSWER200 bp to 50 kb pulse field gel electrophoresis - ANSWERused to separate large DNA up to 10,000 kb repeated changes of electrical current direction TAE buffer - ANSWERtris acetate with EDTA req. re-circulation faster than TBE TBE buffer - ANSWERtris borate wiith EDTA smaller fragments higher buffering capacity methods of gel staining: - ANSWER1. EtBr 2. silver staining 3. SYBR green 4. Methylene Blue SYBR Green I and II - ANSWERI= ds DNA II= ss and RNA nitrocellulose membranes - ANSWERfragile, brittle NOT for re-probing intense color Nylon membrane - ANSWERstrong, durable can strip and re-probe can be charged NOT for Western Blotting Southern Blot - ANSWERDNA Western Blot - ANSWERprotein Northern Blot - ANSWERRNA steps of pre-transfer for membranes - ANSWER1. de-purinate DNA to reduce the size of fragments to transfer 2. denature DNA 3. neutralize to equlibrate gel and membrane capillary transfer to membrane - ANSWERtransfer buffer drawn up through a reservoir and passes through gel into a stack of paper towels weighted; DNA moves from gel with movement of buffer does NOT work forpolyacrylamide gels dot blots - ANSWERbind DNA or RNA to membrane without first separating the components by electrophoresis