BIO 340 LATEST TIPS ANSWERS AND QUESTIONS
SET A+
✔✔reaction ingredients - ✔✔1. DNA template with region to be copied (amplified)
2. dkeoxynuceotide triphosphate= dNTPs (dCTP, dTTP, dATP, dGTP)
3. polymerase (usually Taq)
4. aqueous buffer (including MgCl₂)
5.DNA primers
6. fluorescently labeled dideoxynucleotide triphosphate= ddNTPs
✔✔Which of the following is the correct sequence of steps in a typical PCR sequence -
✔✔denaturation, primer annealing, primer extension
✔✔Denaturation - ✔✔reaction is heated to approx. 95 degrees C to separate DNA
✔✔Primer annealing - ✔✔reaction is heating cooled to 45-68 degrees C to allow
primers to bind to template strand
✔✔Primer extension - ✔✔reaction is heated to approx. 72 degrees C for primer
extension (synthesis of target DNA)
✔✔Reaction components - ✔✔primer sequence
concentration of MgCl₂
✔✔Reaction conditions - ✔✔annealing temperature (=5 degrees C lower than Tm)
extension time (=1 minute per kb)
✔✔Sequencing - ✔✔-Sanger or dideoxy sequencing
-next generation or sequencing by synthesis
-long read, single molecule, third generation sequencing
✔✔illumina sequencing - ✔✔-double stranded, blunt-end DNA fragments are generated
by shearing
- a double-stranded primer pair is ligated
,-DNA is denatured and amplified
✔✔A loss of function mutation to a haplosufficient gene produces a - ✔✔recessive
mutation
-loss-of-function mutations are RECESSIVE if the normal allele is haplosufficient
✔✔A loss of function mutation to a haploINsufficient gene produces a - ✔✔dominant
mutation
-loss-of-function mutations are DOMINANT if the normal allele is haploINsufficient
✔✔Incomplete Dominance - ✔✔the phenotype of the heterozygote is intermediate
between those of the two homozygotes, on some quantitative scale (color, size etc.)
✔✔Co-dominance - ✔✔the phenotype of both alleles is fully expressed in the
heterozygote
✔✔Gene Interaction - ✔✔occurs when two or more genes affect the same phenotype,
by influencing a common pathway
✔✔*if a dihybrid produces FEWER than 4 phenotypes, then you have evidence that the
two genes are interacting* - ✔✔
✔✔In black-eyed peas, flowers are purple due to the presence of anthocyanin. Two
different genes (P and R) produce enzymes that can convert a while precursor into
anthocyanin. Each gene has a recessive, loss-of-fucntion allele (p and r). What are the
phenotype ratios in a dihybrid cross (PpRr x PpRr) - ✔✔15 purple: 1 white
✔✔Mutation - ✔✔any heritable change to DNA sequence
- some positive
-some negative
-MOST are neutral
✔✔point mutation - ✔✔single or few nucleotides change (can be substitution, insertion,
or deletion)
✔✔substitution mutation - ✔✔change in one nucleotide (can be transition or
transversion)
✔✔transition mutation - ✔✔replacement with the same base type (purine to purine or
pyrimidine to pyrimidine)
✔✔transversion mutation - ✔✔replacement with opposite base type (purine to
pyrimidine or pyrimidine to purine)
, ✔✔Which of the following types of mutation is a transition - ✔✔A-->G
-can't be purine to purine or pyrimidine to pyrimidine)
✔✔Which of the following types of mutation is a transvehrsion? - ✔✔C-->G
-can't be purine to pyrimidine or pyrimidine to purine)
✔✔Silent mutation - ✔✔does not change protein sequence
✔✔missense mutation - ✔✔results in one amino acid change in a protein sequence
✔✔nonsense mutation - ✔✔results in truncated protein sequence due to stop codon
✔✔frameshift mutation - ✔✔results in many amino acids changes downstream of the
mutation and/or truncated protein
✔✔Which class of point mutation produces a product with the same amino acid
sequence as the wild-type polypeptide? - ✔✔silent
-silent mutation are nucleotides substitutions that result in no change in the polypeptide
sequence encoded by the mutant allele
✔✔Which class of point mutation essentially always results in a mutant phenotype? -
✔✔frameshift
-frameshift mutations alter the translational reading frame so that the entire amino acid
sequence is altered beyond the sire of the mutation (nucleotide insertion or deletion)
✔✔what caused spontaneous mutations? - ✔✔-nucleotides structural changes
-(tautomerism)
-nucleotide structural damage
- (lesions)
-DNA strand slippage
-(replication error)
✔✔*nucleotide structural isomerism and sessions occur many thousands times in each
cells every 24 hours* - ✔✔-however, most events are quickly revered or repaired and
do not result in DNA mutation
✔✔tautomers
*most common form of replication error* - ✔✔when nucleotide bases can occasionally
convert between alternative structures
✔✔radiation induced damage (usually uv) - ✔✔photo reactive (=photoreactive repair)
UV repair (by nucleotide excision)
✔✔mismatch repair - ✔✔-mismatch repair in E. coli
SET A+
✔✔reaction ingredients - ✔✔1. DNA template with region to be copied (amplified)
2. dkeoxynuceotide triphosphate= dNTPs (dCTP, dTTP, dATP, dGTP)
3. polymerase (usually Taq)
4. aqueous buffer (including MgCl₂)
5.DNA primers
6. fluorescently labeled dideoxynucleotide triphosphate= ddNTPs
✔✔Which of the following is the correct sequence of steps in a typical PCR sequence -
✔✔denaturation, primer annealing, primer extension
✔✔Denaturation - ✔✔reaction is heated to approx. 95 degrees C to separate DNA
✔✔Primer annealing - ✔✔reaction is heating cooled to 45-68 degrees C to allow
primers to bind to template strand
✔✔Primer extension - ✔✔reaction is heated to approx. 72 degrees C for primer
extension (synthesis of target DNA)
✔✔Reaction components - ✔✔primer sequence
concentration of MgCl₂
✔✔Reaction conditions - ✔✔annealing temperature (=5 degrees C lower than Tm)
extension time (=1 minute per kb)
✔✔Sequencing - ✔✔-Sanger or dideoxy sequencing
-next generation or sequencing by synthesis
-long read, single molecule, third generation sequencing
✔✔illumina sequencing - ✔✔-double stranded, blunt-end DNA fragments are generated
by shearing
- a double-stranded primer pair is ligated
,-DNA is denatured and amplified
✔✔A loss of function mutation to a haplosufficient gene produces a - ✔✔recessive
mutation
-loss-of-function mutations are RECESSIVE if the normal allele is haplosufficient
✔✔A loss of function mutation to a haploINsufficient gene produces a - ✔✔dominant
mutation
-loss-of-function mutations are DOMINANT if the normal allele is haploINsufficient
✔✔Incomplete Dominance - ✔✔the phenotype of the heterozygote is intermediate
between those of the two homozygotes, on some quantitative scale (color, size etc.)
✔✔Co-dominance - ✔✔the phenotype of both alleles is fully expressed in the
heterozygote
✔✔Gene Interaction - ✔✔occurs when two or more genes affect the same phenotype,
by influencing a common pathway
✔✔*if a dihybrid produces FEWER than 4 phenotypes, then you have evidence that the
two genes are interacting* - ✔✔
✔✔In black-eyed peas, flowers are purple due to the presence of anthocyanin. Two
different genes (P and R) produce enzymes that can convert a while precursor into
anthocyanin. Each gene has a recessive, loss-of-fucntion allele (p and r). What are the
phenotype ratios in a dihybrid cross (PpRr x PpRr) - ✔✔15 purple: 1 white
✔✔Mutation - ✔✔any heritable change to DNA sequence
- some positive
-some negative
-MOST are neutral
✔✔point mutation - ✔✔single or few nucleotides change (can be substitution, insertion,
or deletion)
✔✔substitution mutation - ✔✔change in one nucleotide (can be transition or
transversion)
✔✔transition mutation - ✔✔replacement with the same base type (purine to purine or
pyrimidine to pyrimidine)
✔✔transversion mutation - ✔✔replacement with opposite base type (purine to
pyrimidine or pyrimidine to purine)
, ✔✔Which of the following types of mutation is a transition - ✔✔A-->G
-can't be purine to purine or pyrimidine to pyrimidine)
✔✔Which of the following types of mutation is a transvehrsion? - ✔✔C-->G
-can't be purine to pyrimidine or pyrimidine to purine)
✔✔Silent mutation - ✔✔does not change protein sequence
✔✔missense mutation - ✔✔results in one amino acid change in a protein sequence
✔✔nonsense mutation - ✔✔results in truncated protein sequence due to stop codon
✔✔frameshift mutation - ✔✔results in many amino acids changes downstream of the
mutation and/or truncated protein
✔✔Which class of point mutation produces a product with the same amino acid
sequence as the wild-type polypeptide? - ✔✔silent
-silent mutation are nucleotides substitutions that result in no change in the polypeptide
sequence encoded by the mutant allele
✔✔Which class of point mutation essentially always results in a mutant phenotype? -
✔✔frameshift
-frameshift mutations alter the translational reading frame so that the entire amino acid
sequence is altered beyond the sire of the mutation (nucleotide insertion or deletion)
✔✔what caused spontaneous mutations? - ✔✔-nucleotides structural changes
-(tautomerism)
-nucleotide structural damage
- (lesions)
-DNA strand slippage
-(replication error)
✔✔*nucleotide structural isomerism and sessions occur many thousands times in each
cells every 24 hours* - ✔✔-however, most events are quickly revered or repaired and
do not result in DNA mutation
✔✔tautomers
*most common form of replication error* - ✔✔when nucleotide bases can occasionally
convert between alternative structures
✔✔radiation induced damage (usually uv) - ✔✔photo reactive (=photoreactive repair)
UV repair (by nucleotide excision)
✔✔mismatch repair - ✔✔-mismatch repair in E. coli
