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(MB) ASCP Study Exam Questions
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What is metagenome? - Sampling the genome sequence of a community of organisms. Polymorphism involved for Irinotecan? - UGT1A NGS related to deletions and insertions, coverage, depth? - -Paired-end reads for detection of indels -Coverage is how much of your target has been read -Depth is how many times a unique position has been covered Polymerase for lagging strand? - Euk - DNA Pol Alpha Pro - DNA Pol I Polymerase for leading strand? - Euk - DNA Pol Delta and DNA Pol Epsilon Pro - DNA Pol III Polymerase used in LAMP? - -Loop-mediated Isothermal Amplification utilizes Bst Polymerase -Has helicase activity and activity at annealing/elongating temps Highest resistance gene in enterococcus? - vanA has the highest resistance in enterococcus Resistance in mecA gene in MRSA codes for? - mecA responsible for antibiotic resistance in S. aureus (MRSA), and codes for PBP2a, which has low affinity for beta-lactam antibiotics like penicillin. Important in gene expression, defense in eukaryotes? - Person guessed mRNA - gene-expression and codes for proteins that bind to DNA to promote/inhibit expression. Donor/Recipient - what's informative loci? - No matching alleles is informative, you can tell D from R; non-informative would be both D&R matching. Bond between DNA and histones? - Ionic - DNA is negative and histones are positive Amino Acids that make up histones? - Arginine and Lysine - positively charged amino acids
Multiplex modifications for low yield? - Person guessed KCl. Increasing Mg2+ could increase yield. Decreasing stringency increases yield, but increases chances of mispriming PCR troubleshooting - Increase Temp --> Increase Stringency Increase Probe Length --> Increase Stringency Increase Probe Complexity --> Increase Stringency Lower Salt Conc. --> Increase Stringency Remove Mg2+ --> Increase Stringency Less Time --> Increase Stringency Add Formamide --> Lowers Stringency Add Bovine Serum --> Lowers Stringency Increase Probe Conc. --> Lowers Stringency How does an amplification control help? - Differentiates a true negative from a false negative Amplification controls have a target that should always amplify, even if your sample does not amplify Best molecular assay for Cystic Fibrosis? - CF is caused by a 3b deletion: delF RFLP, PCR and Southern Blot are used. What to use FISH for: - PWS/AS can be detected with FISH Gaucher - Sequencing or enzyme activity screening FXS - RFLP, PCR, SSCP TS - Sequencing or enzyme activity screening How often should a competency be given/done for someone in a complex lab? - Annually Important in Final Lab Report in CLIA? - Lab director signature? Neurotoxin in gel components? - Ethidium Bromide What phenomenon is observed in EtBr in gel electrophoresis? - Products slowed because EtBr alters DNA shape, charge, flexibility Heteroduplex - dsDNA where the complement strands are not completely complementary
Individualized Quality Control Plan. Allows labs to develop their own QC for their unique procedures. SNPs in human genome - 10,000,000 = 10^ Nested PCR question - Most sensitive and specific PCR. Requires 2 sets of primers, 1 pair for two subsequent amps. Couple of primer design questions like pick forward/reverse primers - Primers should be: 20-30 bases long, flanking the target, end in G and/or C on 3' end 40-60% GC content. Tm with a difference of 0-2°C and in the range of 60-65°C What has a Heavy cell T-Cell receptor rearrangement? - Sezary Syndrome CYP2D6 metabolizes what? - Codeine Glassware to rid of RNA? Baking or autoclaving or DEPC? - DEPC inactivates RNAse How mRNA lifespan is lengthened? - poly A tail or 5' methyl cap What gel is used for a linear DNA 200-250 base pairs?
What would be a better sample to extract DNA from formalin-fixed paraffin embedded tissue or 5 year old refrigerated cord blood? - FFPE tissue because the 5 year old cord-blood was refrigerated and not frozen How long until a technologist is required to update their CE's? - 1 year Which would be considered a heteroduplex?
Every time the chromosome is copied it would become shorter if not for what enzyme? - Telomere Transferase You received a 9ng DNA sample and have to make 50ul dilution of this sample at 900pg. How many ul of stock do you need? - Convert 9ng to pg 1ng = 1000pg, so 9ng is equal to 9000pg V1 (amt stock) = V2C 9000pg x ?=50 ul x 900pg answer is 5 A patient is being tested for leukemia and a fusion appears on the patient sample, the positive control and the negative control.( written just like that) What should be done next? Recollect sample and run again, use patient's own RNA, or test for gene rearrangements by Southern blot - test for gene rearrangements by Southern blot How many chromosomes are found in Interphase? - 23 pairs = 46 chromosomes (22 autosomes + 1 sex chromosome) The doctor sent you a patient specimen and two family members' specimens. He wants the patient specimen run for a mutation. If the patient has a mutation , run the family members specimens. If the patient specimen is negative , don't run the family specimens. Do you: Accession all three specimens and run all three specimens and only report the family results if the patient is positive, OR do you accession only the patient specimen , run it and hold the family member specimens until you know the patient results. Or do you accession all three and only run the patient , then decide if to run the family specimens. - Accession all three and only run the patient , then decide if to run the family specimens. The throat swab and a nasopharyngeal specimen are to you on an 11 year old girl. DO you: Accession both specimens under one number and run them together, OR do you call and ask which on to run, OR do you give each one a separate accession number and run both - give each one a separate accession number and run both A patient specimen is cut with RE and yields a 150 fragment + 210 fragment. The control is 360. How many cuts did the restriction enzyme make? - -----150------/-----------210--------------- Therefore, the restriction enzyme is making one cut. Which specimen has the most DNA according to molecular weight? High white cell count, blood clot, low white cell count - High White Cell Count