with Verified Answers
Note - ANSWERSFor this quizlet I basically went in order of all of the recorded lectures,
but Dr. B sometimes jumps around / covers topics multiple times, so if there are cards
where you think "Hey wait a minute haven't I already seen a card on this?" The answer
is prob yes so I am sorry in advance for that lol
In the PCR samples we made, each student had 3 samples. Those samples consisted
of: - ANSWERS2 genes of interest and 1 control.
Luciferase is fond in: - ANSWERSFireflies.
Luciferase is the _____ _____ in our experiment: - ANSWERSnegative control.
Is PC2 expressed in planarians? - ANSWERSYes it is expressed.
Since PC2 is expressed in planarians, that would mean that it is the _____ _____ in our
experiment. - ANSWERSpositive control.
The full name for PC2 is: - ANSWERSProhormone Convertase.
Methods for detecting gene expression using DNA could include: - ANSWERSAnalyzing
chromatin status, analyzing occupancy of promoter by RNA Pol II.
Methods for detecting gene expression using RNA could include: - ANSWERSAnalyzing
RNA levels.
What do we use to measure the RNA levels? - ANSWERSNorthern blot.
The two types of Northern Blotting are: - ANSWERShybridization based, or PCR-based.
In a hybridization-based Northern blot, the strands are _____ and the limits are _____-
_____ - ANSWERSsingle, transcriptome-wide.
In PCR-based Northern blotting, the strands are _____ and the limits are _____-_____ -
ANSWERSsingle (RT-qPCR) and the limits are genome-wide (RNAseq).
The methods for detecting gene expression are: - ANSWERSWestern blotting, mass
spectrometry, ribosome profiling.
,Western blotting involves _____ strands and is _____ based - ANSWERSsingle,
antibody.
Mass-spectrometry is _____ wide - ANSWERSproteome.
Ribosome profiling is _____ wide - ANSWERSproteome (predicted).
How do we sequence DNA? - ANSWERSSanger sequencing.
In 1953, the helical structure was published by - ANSWERSWatson and Crick.
Where are the attachment points for the incoming nucleotides in a RNA / DNA strand? -
ANSWERSAt the 3' hydroxy group.
What part of the nucleotide attaches to the 3' hydroxy group? - ANSWERSThe
phosphate group.
What kind of bond is formed between nucleotides? - ANSWERSphosphodiester bond.
a FREE, unincorporated nucleotides has what 3 components? - ANSWERSTHREE
phosphate groups (triphosphate), a deoxyribose sugar, and a nitrogenous base.
In DNA, how does the structure change form a free, unincorporated nucleotide to an
incorporated nucleotide? - ANSWERSThe free, unincorporated nucleotide will lose two
of its phosphate groups while attaching to the growing strand of DNA / RNA.
What are the nitrogenous bases in DNA? - ANSWERSAdenine, Thymine, Guanine,
Cytosine.
What are the nitrogenous bases in RNA? - ANSWERSAdenine, Uracil, Guanine,
Cytosine
What is unique about a dideoxyribonucleotide compared to a deoxyribonucleotide? -
ANSWERSThe dideoxyribonucleotide does not have a hydroxyl group hanging off of its
3' carbon.
Can you attach nucleotides to dideoxyribonucleotides? - ANSWERSNo, you can't.
Why can't you attach nucleotides to dideoxyribonucleotides? - ANSWERSBecause, they
lack a hydroxy group on the 3' prime carbon, which is where the phosphodiester bond
would be formed. No hydroxy group means no bond.
Dideoxyribonucleotides are used for - ANSWERSChain termination chemistry.
What is Sanger sequencing? - ANSWERSA technique to determine the sequence of
DNA, that uses dideoxynucleotides to prevent further extension past a known base.
, How do you read the gel in Sanger sequencing? - ANSWERSYou look at the dark
bands in each column. There is one column for each type of nucleotide. By reading the
sequence of the bands, you can determine the sequence of nucleotides.
When was Sanger sequencing developed? - ANSWERS1977
Eventually, in 1987, the nucleotides were coupled with _____ molecules. -
ANSWERSFluorescence.
With fluorescence Sanger sequencing, a _____ is used to detect the colors and thus
determine the _____ of the DNA. - ANSWERSlaser, sequence.
The reaction mixture for a Sanger sequencing reaction would include: - ANSWERSA
primer, DNA template, polymerase, ddNTP's with fluorochromes.
Step 2 of Sanger sequencing is: - ANSWERSPrimer elongation and chain termination.
This is where the differing sized strands of DNA are made.
In this mixture, you incorporate both _____ and _____ nucletodies? - ANSWERSdeoxy
and dideoxynucleotides.
Why do you include dideoxynucleotides? - ANSWERSRandomly, it will generate DNA
strands that are all different lengths.
Why is a primer needed in Sanger sequencing? - ANSWERSYou need a primer to
begin the sequence of the complimentary strand. The primer will attach to the template
strand, then polymerase will begin sequencing.
In step 3 of Sanger sequencing: - ANSWERSCapillary gel undergoes a gel
electrophoresis to separate those daggone nucleotides that are different lengths.
In step 4 of the Sanger sequencing reaction - ANSWERSThe laser will detect those
fluorochromes and can comprise a sequence based on the order of them mfkn colors
#yuh.
What is the name of the graph that represents the sequence in a Sanger sequence
reaction? - ANSWERSElectropherogram.
In the electropherogram, sometimes you may get peaks that are weak. If you got some
weak peaks (that rhymes) what could you do to verify the sequence of your DNA? -
ANSWERSYou could synthesize both DNA strands, so the template strand and the
complimentary strand, to double check the peaks on both strands and see if those weak
signals match up.