ASCP Molecular Biology Certification Exam
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Terms in this set (1434)
One carbon ring
Pyrimidine
Cytosine, Thymine, Uracil
What is the function of Carries genetic info out of nucleus
mRNA? Transcript translated to protein
What is the function of Carries aa to ribosome
tRNA? Anticodon pairs with codon on mRNA strand
part of ribosome structure
What is the function of
most abundant RNA
rRNA?
coordinated coupling of tRNA to mRNA codons
Product of pathway is noncompetitive inhibitor
Feedback inhibition Binds to allosteric site to slow down rxn b/c too much
product
Restriction enzymes
Cleaves phoshpodiester bonds w/i poly-nt chain
Endonucleases (Prok)
Recognition site is palindromic sequence
Types I-V
ORI sites nt sequence where replication is initiated
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Induces ss breaks
Remove DNA supercoils during TXN and DNA
Topoisomerase I replication; for strand breakage during recombination;
for chr condensation; and to disentangle intertwined
DNA during mitosis
cuts both strands of one DNA double helix, passes
topoisomerase II another unbroken DNA helix through it, and then
reanneals the cut strands
Unwinds supercoiling caused by unwinding at the rep
Gyrase (topoisomerase II)
fork by introducing DSBs
Helicase Breaks H-bonds of double helix at the replication fork
single-strand DNA binding Binds ssDNA and prevents it from re-annealing during
proteins (SSBPs) TXN, replication, repair, and recombination
Repeat sequence (TTAGGG) at the ends of chr,
Telomeres
protect chr from degradation
DNA dependent RNApol
RNA polymerase Transcribes DNA template to RNA (3'-->5'; anti-
parallel)
Complex of snRNPs
Splicesomes Removes introns from pre-mRNA and splices exons
together
Short regions of DNA that bind proteins (TXN factors)
Enhancers
that enhance TXN of a gene
enzyme that transcribes RNA to cDNA (lacks introns)
Reverse transcriptase
RNA --> RNA:DNA --> cDNA (dsDNA)
sections of DNA that begin with start codons and end
with stop codons
Open Reading Frame
DNA: 5' --> 3'
(ORF)
transcription: 3' --> 5' DNA --> RNA (promoter)
translation: 5' --> 3' mRNA
Measures amount of light absorbed
Spectrophotometer
Quantitative measurement of [DNA/RNA]
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At what wavelength does 260 nm
DNA and RNA absorb?
At what wavelength does 280 nm
protein absorb?
1. Lyse
2. Add phenol/ chloroform > vortex/spin
3. Transfer aqueous layer (top) to new tube
4. Add chloroform:IAA (removes phenol) > vortex/spin
Organic isolation method
5. Transfer aqueous layer to new tube
6. Add NaOAc and EtOH > vortex/spin
7. Decant
8. Resuspend
What does the incubation Allows formation of ds molecules
step in hybridization do?
Formamide acts as a denaturing agent
__________ in a hybridization.
reverse hybridization assay using sequence-specific
Line Probe Assay (LiPA) oligonucleotide probes (reverse SSOP)
multi-parameter testing --> single strip
Used for known gene, unknown mut
Single-Stranded
Mutation screening
Conformational
Short PCR products form 3D conformation when
Polymorphism Ananlysis
cooled --> muts have different conformation than WT
(SSCP)
Non-denaturing PAGE, muts migrate different than WT
How does EtBr cause Intercalates into the double helix
DNA to fluoresce? Absorbs UV ~300 nm, emits ~600 nm
1. culture
2. embed pellet in agarose plug
Pulse Field Gel
3. treat w/ lysozyme (cell lysis)
Electrophoresis steps
4. proteinase K
5. gel
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What are the 3 steps of Denature 90-96C
PCR and their Anneal 50-70C
temperatures? Extension 68-75C
1. RE digest
2. Electrophorese and purify fragment of interest
3. Denature and incubate w/ sodium bisulfate (turns
Bisulfite DNA
C>U, methylated C is unchanged)
sequencing/Methylation
4. clean, ppt, and resuspend
specific
5. PCR --> sequence
6. Compare treated vs untreated, note where CG are
not changed to TA
1. Hybridize oligo-T7P primer to target seq
2. RT/RNase H
NASBA steps
3. Hybridize with target-specific oligo primer (P2)
4. RNA transcript of T7 RNA pol
Do you have to know the Yes, in order to design primers
gene sequence in order to You do NOT need to know the mutation
do DNA sequencing?
Divided into 4 samples (ddA, T, G, C)
Sanger sequencing Label with radioactive/dye oligo at 3' end
method Mix with taq, dNTPs, ddNTP and incubate
run on gel --> frags will terminate at different lengths
Fluorescent in situ Uses fluorescent probes to detect DNA sequences on
hybridization (FISH) chr
Dual fusion: 2 probes flank the breakpoint at both t
locations
What types of probes are CEN probes: centromeric probes bind to repetitive
used for FISH? alpha satellite sequences
Telomeric probes
Whole chr paints
What is the wavelength for 320 nm
background in
spectrophotometery?
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