ASCP Molecular Biology Certification Exam 2024
Pyrimidine - Answer ️️ -One carbon ring
Cytosine, Thymine, Uracil
Purine - Answer ️️ -Two carbon rings
Adenine, Guanine
How are nucleotides joined together? - Answer ️️ -Condensation to form phosphodiester bond
What is the function of...
ASCP Molecular Biology Certification Exam 2024 Pyrimidine - Answer ✔️✔️-One carbon ring Cytosine, Thymine, Uracil Purine - Answer ✔️✔️-Two carbon rings Adenine, Guanine How are nucleotides joined together? - Answer ✔️✔️-Condensation to form phosphodiester bond What is the function of mRNA? - Answer ✔️✔️-Carries genetic info out of nucleus Transcript translated to protein What is the function of tRNA? - Answer ✔️✔️-Carries aa to ribosome Anticodon pairs with codon on mRNA strand What is the function of rRNA? - Answer ✔️✔️-part of ribosome structure most abundant RNA coordinated coupling of tRNA to mRNA codons Feedback inhibition - Answer ✔️✔️-Product of pathway is noncompetitive inhibitor Binds to allosteric site to slow down rxn b/c too much product Exonucleases - Answer ✔️✔️-Degrades nucleic aci ds by removing one terminal nt at a time Cleaves phosphodiester bond at end of chain 5' --> 3' and 3' --> 5' Endonucleases (Prok) - Answer ✔️✔️-Restriction enzymes Cleaves phoshpodiester bonds w/i poly -nt chain Recognition site is palindromic sequence Types I-V ORI sites - Answer ✔️✔️-nt sequence where replication is initiated Topoisomerase I - Answer ✔️✔️-Induces ss breaks Remove DNA supercoils during TXN and DNA replication; for strand breakage during recombination; for chr condensation; and to disenta ngle intertwined DNA during mitosis topoisomerase II - Answer ✔️✔️-cuts both strands of one DNA double helix, passes another unbroken DNA helix through it, and then reanneals the cut strands Gyrase (topoisomerase II) - Answer ✔️✔️-Unwinds supercoiling ca used by unwinding at the rep fork by introducing DSBs Helicase - Answer ✔️✔️-Breaks H -bonds of double helix at the replication fork Primase - Answer ✔️✔️-DNApol α (DNA dep RNA pol) adds short segments of complementary RNA to ssDNA template (primers), se rves as starting points for replication single -strand DNA binding proteins (SSBPs) - Answer ✔️✔️-Binds ssDNA and prevents it from re-annealing during TXN, replication, repair, and recombination Okazaki fragments - Answer ✔️✔️-Short fragments of DNA synth esized by DNApol δ using the lagging strand (3' ->5') as a template Ligase - Answer ✔️✔️-Closes gaps in DNA Catalyzes phosphodiester bond between 3'OH and 5'P What are the steps in DNA replication? - Answer ✔️✔️-1. Initiate 2. Elongate 3. Terminate Telom eres - Answer ✔️✔️-Repeat sequence (TTAGGG) at the ends of chr, protect chr from degradation RNA polymerase - Answer ✔️✔️-DNA dependent RNApol Transcribes DNA template to RNA (3' -->5'; anti -parallel) Splicesomes - Answer ✔️✔️-Complex of snRNPs Removes i ntrons from pre -mRNA and splices exons together Enhancers - Answer ✔️✔️-Short regions of DNA that bind proteins (TXN factors) that enhance TXN of a gene Poly-A tail - Answer ✔️✔️-Prevents mRNA from being degraded in cytoplasm 100-250 A's at 3' end 5' cap - Answer ✔️✔️-5'-5' pyrophosphate bridge to a methylated G added to 5' end of a mRNA Protects against degradation and as a recognition signal for TLN apparatus aminoacyl tRNA - Answer ✔️✔️-tRNAs that carry amino acids Ribosomes - Answer ✔️✔️-Where TLN occurs Prok: 30s and 50s Euk: 40s and 60s Catalyzes peptide bond between a.a.'s What is the path of a tRNA in a ribosome? - Answer ✔️✔️-Acceptor > Peptidyl > Exit How is translation initiated? - Answer ✔️✔️-small rRNA (40S) subunit binds mRNA and scans for start codon (AUG) Met-tRNA is brought to the P site Large rRNA (60S) subunit binds How is translation terminated? - Answer ✔️✔️-Occurs when stop codon enters A site Release factor recognizes stop codon, hydrolyzes ester bond with P site, releasing aa c hain Reverse transcriptase - Answer ✔️✔️-enzyme that transcribes RNA to cDNA (lacks introns) RNA --> RNA:DNA --> cDNA (dsDNA) Pleiotrophy - Answer ✔️✔️-a single gene controls the expression of many phenotypic traits ie Sickle Cell Anemia cDNA - Answer ✔️✔️-intron free complementary DNA can be inserted into a plasmid Vector - Answer ✔️✔️-helps carry DNA into cell ie plasmids, virus Open Reading Frame (ORF) - Answer ✔️✔️-sections of DNA that begin with start codons and end with stop codons DNA: 5' --> 3' transcription: 3' --> 5' DNA --> RNA (promoter) translation: 5' --> 3' mRNA Spectrophotometer - Answer ✔️✔️-Measures amount of light absorbed Quantitative measurement of [DNA/RNA] At what wavelength does DNA and RNA absorb? - Answer ✔️✔️-260 nm At wha t wavelength does protein absorb? - Answer ✔️✔️-280 nm Organic isolation method - Answer ✔️✔️-1. Lyse 2. Add phenol/ chloroform > vortex/spin 3. Transfer aqueous layer (top) to new tube 4. Add chloroform:IAA (removes phenol) > vortex/spin 5. Transfer aqu eous layer to new tube 6. Add NaOAc and EtOH > vortex/spin 7. Decant 8. Resuspend How do you inactivate RNases? - Answer ✔️✔️-200C for 2 hrs 30 min in 1M NaOH or quanidinum isothiocyanate Hybridization - Answer ✔️✔️-2 ssDNA molecules of comp base sequenc e can form a ds hybrid (duplex) What does the incubation step in hybridization do? - Answer ✔️✔️-Allows formation of ds molecules Blocking DNA (Hybridization) - Answer ✔️✔️-minimizes probe binding to nonspecific sequence ie salmon sperm DNA, Human LINE -1 Blocking Proteins (Hybridization) - Answer ✔️✔️-minimize nonspecific binding of probe to membrane ie casein (milk), Denhardt's sol Stringency - Answer ✔️✔️-conditions of hybridization that control the specificity of binding of the probe to the target se quence How can you increase strigency in a hybridization? - Answer ✔️✔️-decrease [salt] increase [formamide] increase temp Formamide acts as a __________ in a hybridization. - Answer ✔️✔️-denaturing agent Line Probe Assay (LiPA) - Answer ✔️✔️-reverse hy bridization assay using sequence -specific oligonucleotide probes (reverse SSOP) multi -parameter testing --> single strip
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