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ASCP Molecular Biology Certification Exam 2024 with 100% correct answers

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Pyrimidine correct answersOne carbon ring Cytosine, Thymine, Uracil What is the function of mRNA? correct answersCarries genetic info out of nucleus Transcript translated to protein What is the function of tRNA? correct answersCarries aa to ribosome Anticodon pairs with codon on mRNA stran...

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ASCP Molecular Biology Certification
Exam

Pyrimidine correct answersOne carbon ring

Cytosine, Thymine, Uracil



What is the function of mRNA? correct answersCarries genetic info out of nucleus

Transcript translated to protein



What is the function of tRNA? correct answersCarries aa to ribosome

Anticodon pairs with codon on mRNA strand



What is the function of rRNA? correct answerspart of ribosome structure

most abundant RNA

coordinated coupling of tRNA to mRNA codons



Feedback inhibition correct answersProduct of pathway is noncompetitive inhibitor

Binds to allosteric site to slow down rxn b/c too much product



Endonucleases (Prok) correct answersRestriction enzymes

Cleaves phoshpodiester bonds w/i poly-nt chain

Recognition site is palindromic sequence

Types I-V



ORI sites correct answersnt sequence where replication is initiated



Topoisomerase I correct answersInduces ss breaks

,Remove DNA supercoils during TXN and DNA replication; for strand breakage during recombination; for
chr condensation; and to disentangle intertwined DNA during mitosis



topoisomerase II correct answerscuts both strands of one DNA double helix, passes another unbroken
DNA helix through it, and then reanneals the cut strands



Gyrase (topoisomerase II) correct answersUnwinds supercoiling caused by unwinding at the rep fork by
introducing DSBs



Helicase correct answersBreaks H-bonds of double helix at the replication fork



single-strand DNA binding proteins (SSBPs) correct answersBinds ssDNA and prevents it from re-
annealing during TXN, replication, repair, and recombination



Telomeres correct answersRepeat sequence (TTAGGG) at the ends of chr, protect chr from degradation



RNA polymerase correct answersDNA dependent RNApol

Transcribes DNA template to RNA (3'-->5'; anti-parallel)



Splicesomes correct answersComplex of snRNPs

Removes introns from pre-mRNA and splices exons together



Enhancers correct answersShort regions of DNA that bind proteins (TXN factors) that enhance TXN of a
gene



Reverse transcriptase correct answersenzyme that transcribes RNA to cDNA (lacks introns)

RNA --> RNA:DNA --> cDNA (dsDNA)



Open Reading Frame (ORF) correct answerssections 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 correct answersMeasures amount of light absorbed

Quantitative measurement of [DNA/RNA]



At what wavelength does DNA and RNA absorb? correct answers260 nm



At what wavelength does protein absorb? correct answers280 nm



Organic isolation method correct answers1. 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 aqueous layer to new tube

6. Add NaOAc and EtOH > vortex/spin

7. Decant

8. Resuspend



What does the incubation step in hybridization do? correct answersAllows formation of ds molecules



Formamide acts as a __________ in a hybridization. correct answersdenaturing agent



Line Probe Assay (LiPA) correct answersreverse hybridization assay using sequence-specific
oligonucleotide probes (reverse SSOP)

multi-parameter testing --> single strip



Single-Stranded Conformational Polymorphism Ananlysis (SSCP) correct answersUsed for known gene,
unknown mut

, Mutation screening

Short PCR products form 3D conformation when cooled --> muts have different conformation than WT

Non-denaturing PAGE, muts migrate different than WT



How does EtBr cause DNA to fluoresce? correct answersIntercalates into the double helix

Absorbs UV ~300 nm, emits ~600 nm



Pulse Field Gel Electrophoresis steps correct answers1. culture

2. embed pellet in agarose plug

3. treat w/ lysozyme (cell lysis)

4. proteinase K

5. gel



What are the 3 steps of PCR and their temperatures? correct answersDenature 90-96C

Anneal 50-70C

Extension 68-75C



Bisulfite DNA sequencing/Methylation specific correct answers1. RE digest

2. Electrophorese and purify fragment of interest

3. Denature and incubate w/ sodium bisulfate (turns C>U, methylated C is unchanged)

4. clean, ppt, and resuspend

5. PCR --> sequence

6. Compare treated vs untreated, note where CG are not changed to TA



NASBA steps correct answers1. Hybridize oligo-T7P primer to target seq

2. RT/RNase H

3. Hybridize with target-specific oligo primer (P2)

4. RNA transcript of T7 RNA pol

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