exam 1 Flashcards
true or false? you need an electroporation cuvette for heat shock transformation of E.coli
false
Which of the following are parts of an adaptor used for preparation and sequencing of genomic DNA fragments
A) a sequence that anneals to an oligoneculeotide primer
B) a sequence that anneals to an sequencing primer to prime the sequencing reaction
C) a unique barcode sequence used to tag a genomic library
all of the above
TA cloning is a PCR based cloning true or false
True
you extracted RNA from a drought stressed plant leaf. you are interested in cloning only drought stressed cDNAs. what type of primer will you use at the very beginning after the sythesis of mRNA?
Gene specific primer
T4 polynucleotide kinase is used in Dr. Sanger’s method of DNA sequencing. true or false
false
in crisper system for use in biotech can the sgRNA and Cas9 gene be cloned in the same plasmid
yes
neoschizomers are restriction enzymes that
can bind at the same recognition site but cleave at differnt sites
a ddNTP has
a H group at the 3’ carbon
what is true about the oxidation step during primer synthesis
this step makes the bonds between the neucleotides stronger
at the time of the gene synthesis by primers how can you seal the nicks in the strands of DNA
by adding DNA ligase
is a chemical blocking group attatched to the 3’ carbon of the sugar moiety during pyrosequencing ?
no
what is true about capping during primer sythesis
capping is done to prevent prmers differing in lengths
luciferase+_____–> light
ATP
what is pyrosequencing?
a cleaved beta and gammma phosphate from dNTp molecule
MALDI-TOF analysis of protein molecules are represented by ?
mass to charge ratio
you want to repair the ends of DNA repair means the addition of nucleotides what type of enzyme do you use?
DNA polymerase
in colony immunoassay, first cells are lysed to release___ bound on the membrane
protein
you are identifying disease biomarkers with human protein microarray. what should be spotted onto the slide?
human proteins
what enzyme is used to “paste” your gene of intrest fragment into a plasmid cloning vector?
ligase
is DMT group part of a phosphoramidite?
yes
you are using “sandwich’ style protein microarray to study post translational modifications. should the proteins be labeled with flourescent tags?
no
you can analyze millions of transcripts using RNAseq? true or false
true
during primer synthesis how can you remove a DMT group?
by addition of TCA
in CRISPR invadiong foreign DNA have a sequence similarity with____
Crispr loci
which sequencing method has the lowest accuracy
Single molecule sequencing
the ccdB gene used in recombination cloning (gateway cloning) is a
gene that produces toxic compound
you extracted DNA then what is the first step in shotgun sequencing?
break genomic DNA
shine dalgarno sequence is
a ribosomal binding site in prokaryotes
in crisper system the invading DNA must have a
PAM sequence
in qPCR. if sample A has a Ct value of 20 and sample B has a Ct value of 40 in the same qPCR experiment can you canclude sample A has more DNA than sample B
yes
a kinase
transfers a phosphate group to a DNA
in blue white colony screening
LacZ gene encodes for beta galactosidase in the presense of X-Gal
activation and coupling during primer synthesis are mediated by
tetrazole
Dr. venter performed comprehensive construction of DNA libraries from all microorganisms from a region in Saragasso sea. this type fo study is known as
metagenomic study
During cDNA synthesis you extracted total RNA and by mistake added RNAseh instead of reverse transcript. what will happen?
my RNA will be degraded and i will not be able to continue cDNA synthesis
during the primer synthesis the purpose of adding a DMT group to a nucleotide is
to prevent unlinked nucleosides reacting non specifically
you are using a yeast two hybrid system. you have 100 yeast strains containing DNA fragments as prey library. can you mix all prey yeast strains into a tube and have the bait yeast strains mate with the prey
yes
your goal is to synthesize a gene. you synthesized oligonucleotides what should be now the immediate next step
increase temperature to 95*C for denaturation
which of the following enzymes is able to synthesize another strand of DNA from a template DNA molecule
Klenow (DNA) polymerase
the SYBR green dye used in qPCR can bind with both single and double stranded DNA
no
what is true about metagenomic library construction
you need to perform shotgun sequencing
what is detriylation?
the removal of DMT from necleoside during primer synthesis
you can use pUC 19 for blue white colony screaning
true
during synthesis of oligonucleotides the amino group is removed all nucleotides except
T
find a restriction enzyme producing blunt ended DNA fragment
Pvull
pyrophosphate +______ =ATP
ATP sulfrylase
gel is needed for qPCR
false
in crispr the components of single guide RNA is crRNA and _______
tracrRNA
in tandem affinity purification procedure a plasmid DNA is used with two tags. which sequence is inserted between the two tags?
protease clevage site
a 2D gel elecrephoresis system separates protein on the bands of elecgtric gradient an ____ gradien
pH
what site/genetic element is an essential component of phage DNA to construct a phage library
COS
what is the realistic proportion of mRNA in total RNA sample?
3%
what is the role of SEC complex in protein transport in gram positive bacteria
creates a passage thx which entire protein and signal peptide passes
what is the prinbrow box
TATAAT
a bacteriophage life cycle goes through lysogeny and lytic cycle after it infects E. coli which is true?
in lysogeny phage DNA gets integrated in bacterial genome and maaintained in bacterial genome
what are the number 5 and 3 in 5’ and 3’ represent when we describe the orientation of DNA
number on the carbon atom of the nucleotide
biotin streptabin conjugate is used to detect numerous biomolecules including nucleic acids
true
all of the antibiotics below inhibit protein sythesis and kill cells which one does not inhibit protein synthesis but still kills the cell
ampicillin
at the time of labeling a probe it is nescessary to boil the probe. why?
boling denatures the probe and only a single stranded denatured probe can hybridize effectively with target DNA
what is southern blot hybridization
the gene of intrest (probe) is labled and total genomic DNA is loaded onto the gel
which enzyme is used to “paste” GOI fragment in a cloning vector
ligase
find a restriction enzyme producing blunt ended DNA fragments
PvuII
Neoshizomers are RE that can»_space;»
bind at the same recognition site but cleave at a differnt site
TATA box
promoter region
RNA polymerase
binds to promoter region in the DNA to initiate transcription
if a sequence of a template strand of a DNA is CAATGA the sequence of mRNA synthesized will be
GUUACU
an mRNA molecule is successfully alternitively spliced to form two functional coding mRNA. alternitive splicing can generate two:
differnt types of mRNA with differnt exons
arrange the following events in transcription and translation
RNA polymerase binds to DNA
ribosome is attatched to
to mRNA
mRNA sythesis
RNA polymerase binds toDNA -> mRNA synthesis-> ribosome is attached to mRNA
and mRNA molecule has a unique feature which is absent in tRNA or rRNA
a poly a tail
in blue white colony screening whats the role of IPTG
it inactivated repressor protein
which of the following is not a part of the trnscription factor complex
TFIIZ
as soon as proteins are synthesized from ribosome-mRNA complex the protein are loaded into the
rough endoplasmic reticulum
what is the disadvantage of pBR322 cloning vector
very few cloning sites
sometimes the functional genetic complementation is used at the time of gene cloning which is true of functional genetic complemntation
you can study gene response for host cell specific enzyme activity, make DNA library, transform host cells with DNA library
all of the above
what is the smallest insert capacity to largest?
plasmids>bacteriophage lambda>BAC>YAC
what does deoxyribose look like
2’H 3’ OH
what is the finction of RNASeH
digest any type of RNA double or single stranded
what is the disadvantage fo shittle vector
they can be lost from the host cell
which one is the worst cloning vector for human genome sequencing
plasmid
which is true of dynabead
have oligodT attached to it
what is the role if kinase
transfer phosphate to donor molecules to substrates through phosphorylation
is it posible for plant to have larger genome than humans
yes statment is true in some case but not all
Which is true about klenow frag.
its a product of E. coli DNA polym
Which is NOT true about pBR322 plasmid vector
has a lac z gene
What’s true about gene cloning by functional complementation technique we studied in class
A. All correct; need normal and defective E. coli strain, need genomic DNA library w. Goi cloned, need to grow
cells on min. Medium
Which enzyme is exclusively used in RNA manipulation but not in DNA manipulation
A. Poly A polymerase
How do you develop EST library?
By random sequencing of cDNAs into a CDNA library
You work for USDA and interested in gene expression in corn plants. You can perform
A. Microarray and northern blot
