Technologist in Molecular Biology, MB(ASCP) board exam
prep 2024/2025 with 100% Correct Answers
Pyrimidine - ✔️✔️-One carbon ring
Cytosine, Thymine, Uracil
Purine - ✔️✔️-Two carbon rings
Adenine, Guanine
How are nucleotides joined together? - ✔️✔️-Condensation to form phosphodiester
bond
What is the function of mRNA? - ✔️✔️-Carries genetic info out of nucleus
Transcript translated to protein
What is the function of tRNA? - ✔️✔️-Carries aa to ribosome
Anticodon pairs with codon on mRNA strand
What is the function of rRNA? - ✔️✔️-part of ribosome structure
most abundant RNA
coordinated coupling of tRNA to mRNA codons
Feedback inhibition - ✔️✔️-Product of pathway is noncompetitive inhibitor
Binds to allosteric site to slow down rxn b/c too much product
Exonucleases - ✔️✔️-Degrades nucleic acids by removing one terminal nt at a time
Cleaves phosphodiester bond at end of chain
5' --> 3' and 3' --> 5'
Endonucleases (Prok) - ✔️✔️-Restriction enzymes
Cleaves phoshpodiester bonds w/i poly-nt chain
Recognition site is palindromic sequence
Types I-V
ORI sites - ✔️✔️-nt sequence where replication is initiated
, Technologist in Molecular Biology, MB(ASCP) board exam
prep 2024/2025 with 100% Correct Answers
Topoisomerase I - ✔️✔️-Induces 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 - ✔️✔️-cuts both strands of one DNA double helix, passes another
unbroken DNA helix through it, and then reanneals the cut strands
Gyrase (topoisomerase II) - ✔️✔️-Unwinds supercoiling caused by unwinding at the
rep fork by introducing DSBs
Helicase - ✔️✔️-Breaks H-bonds of double helix at the replication fork
Primase - ✔️✔️-DNApol α (DNA dep RNA pol)
adds short segments of complementary
RNA to ssDNA template (primers), serves as starting points for replication
single-strand DNA binding proteins (SSBPs) - ✔️✔️-Binds ssDNA and prevents it from
re-annealing during TXN, replication, repair, and recombination
Okazaki fragments - ✔️✔️-Short fragments of DNA synthesized by DNApol δ using the
lagging strand (3'->5') as a template
Ligase - ✔️✔️-Closes gaps in DNA
Catalyzes phosphodiester bond between 3'OH and 5'P
What are the steps in DNA replication? - ✔️✔️-1. Initiate
2. Elongate
3. Terminate
Telomeres - ✔️✔️-Repeat sequence (TTAGGG) at the ends of chr, protect chr from
degradation
RNA polymerase - ✔️✔️-DNA dependent RNApol
, Technologist in Molecular Biology, MB(ASCP) board exam
prep 2024/2025 with 100% Correct Answers
Transcribes DNA template to RNA (3'-->5'; anti-parallel)
Splicesomes - ✔️✔️-Complex of snRNPs
Removes introns from pre-mRNA and splices exons together
Enhancers - ✔️✔️-Short regions of DNA that bind proteins (TXN factors) that enhance
TXN of a gene
Poly-A tail - ✔️✔️-Prevents mRNA from being degraded in cytoplasm
100-250 A's at 3' end
5' cap - ✔️✔️-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 - ✔️✔️-tRNAs that carry amino acids
Ribosomes - ✔️✔️-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? - ✔️✔️-Acceptor > Peptidyl > Exit
How is translation initiated? - ✔️✔️-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? - ✔️✔️-Occurs when stop codon enters A site
Release factor recognizes stop codon, hydrolyzes ester bond with P site, releasing aa
chain
Reverse transcriptase - ✔️✔️-enzyme that transcribes RNA to cDNA (lacks introns)
, Technologist in Molecular Biology, MB(ASCP) board exam
prep 2024/2025 with 100% Correct Answers
RNA --> RNA:DNA --> cDNA (dsDNA)
Pleiotrophy - ✔️✔️-a single gene controls the expression of many phenotypic traits
ie Sickle Cell Anemia
cDNA - ✔️✔️-intron free complementary DNA
can be inserted into a plasmid
Vector - ✔️✔️-helps carry DNA into cell
ie plasmids, virus
Open Reading Frame (ORF) - ✔️✔️-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 - ✔️✔️-Measures amount of light absorbed
Quantitative measurement of [DNA/RNA]
At what wavelength does DNA and RNA absorb? - ✔️✔️-260 nm
At what wavelength does protein absorb? - ✔️✔️-280 nm
Organic isolation method - ✔️✔️-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 aqueous layer to new tube
6. Add NaOAc and EtOH > vortex/spin
7. Decant
8. Resuspend
How do you inactivate RNases? - ✔️✔️-200C for 2 hrs
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