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Questions:
1.
You should note that the thermocycler has a heated lid that covers your tubes. Why do
you think a heated lid is necessary?
a heated lid prevents condensation in the top of the tube.
2.
When designing the oligonucleotides for this experiment, it was determined that the
Tm (melting temperature) of BlaF (oDM0318) and BlaR (oDM0319) was 61
o
C and
64.6
o
C, respectively. This knowledge is important for determining the annealing step?
Explain how you define the annealing temperature and why?
The annealing temperature is about 3-5 degrees below the the TM of the primers which allows
the annealing of the DNA oligonucleotide primers to the single stranded DNA template. This
eventually leads to DNA synthesis.
3.
Looking at reagents contained in Master Mixes, can you define the purpose of each of
those listed below.
a. Deoxynucleotides:
Building blocks from which the DNA polymerase synthesizes a new
strand, dATP, dCTP,dCTP,dTTP
b. Mg
+2
: used by Taq Polymerase to carry synthesis of the new strands
c. Taq polymerase: synthesizes the newly formed strands by adding bases complementary
in A5 prime to three prime direction
d. BlaF/BlaR: these allow each single strand DNA molecule to be read in the/ reverse
direction left to right/right to left
4. The sequence of the oligonucleotides used for the PCR amplification of the Ampicillin
resistance gene (Bla) are:
1.
BlaF (oDM0318): 5’
GAGTATTCAACATTTCCGTGTCG 3’
‐
‐
2.
BlaR (oDM0319): 5’
CCAATGCTTAATCAGTGAGGCACC 3’
‐
‐
The sequence of the ampicillin resistance gene is shown below. Please highlight or underline
where BlaF and BlaR are annealing to the DNA for amplification. Based on this knowledge,
you should be able to predict the size of the amplified DNA fragment you will observe on the
agarose gel that you will run next week. What is the predicted size of the PCR product of the
Bla gene?
PCR fragment size is ____858_____ bp.
Table 5: Data Table for the PCR reaction
1 2 3 4
Questions:
Bands from PCR observed (top to bottom
of gel)
Distance migrated from the well
(mm)
Size of the bands in bp.
1
no bands
0mm
2
two bands
37mm, 64mm
1,000bp,500bp
3
three bands
17mm, 28mm, 38mm
4,000bp, 1,500bp,
1,000bp
4
1.
How does the predicted size of the PCR product correlate with the size you measured
in Table 5?
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Related Questions
questions 23-31
arrow_forward
#41 and #42
arrow_forward
Important steps in sequencing pure proteins include all of these except:
a. Determining the amino acids on the ends of the smaller peptides
b. Determining the amino acids on the ends of the protein
c. Determining the isoionic pH of the protein
d. Determining the amino acid composition
e. Breaking the protein into smaller peptides
It is impossible to sequence a protein if you do not have overlapping sequences to work with.
A. True
B. False
arrow_forward
Based on the following nucleotide structure, answer the following questions:
H2N
N-
Но-Р-о-
N.
OH
a. Is the base present in this nucleotide a purine or a pyrimidine?
b. What is the abbreviation for the base present in this nucleotide? (A, T, C, G, U)
Would the sugar in this nucleotide be found in molecules of DNA or RNA or both?
d. Would the base in this nucleotide be found in molecules of DNA, RNA or both?
arrow_forward
Are the following compounds correct?
These may OR may not be correct. Please look them over and correct them if needed.
Please list what was done incorrectly if applicable.
b. Is the following nucleotide found in DNA?
If not, please explain why.
a. Is the following tRNA correct?
NH2
VALINE
<5'
N.
3'-
0=P-0-CH2
ОН ОН
GUA
a.
b.
DNAA=T
G=C
arrow_forward
On your Scantron,
best possible answer for each question and fill in the corresponding bubble on your Scantron
There are 50 total questions, each worth 3 points.
1. Which structure represents a nucleotide?
A)
CH,O-
-O. 0-
OH
ОН
B)
NH2
CH,
H
H
OH
OH
C)
HO-CH,
OH
H
H
H.
OH
H
H
D)
NH,
0.
CH,
OH
OH
0-P=0
2. Which vitamin is fat soluble?
arrow_forward
nucleotides 2
Which component is not found in a nucleotide?
Select an answer and submit. For keyboard navigation, use the up/down arrow keys to select an answer.
phosphate
b.
base
sulfate
saccharide
arrow_forward
The DNA of bacteriophage lambda has 1.2 x 10^5 nucleotides. How many proteins of molecular weight 40,000 could be coded by this DNA? Assume a molecular weight of 100 for the average amino acid.
arrow_forward
ANALYSIS
1. A viral genome is composed of a double stranded DNA molecule containing 14% T (thymine). Based on this information, what would be the percentage of cytosine?
arrow_forward
1. Draw the two Watson-Crick Bases Pairs and show how G-
C and A-T recognize each other by hydrogen bonding.
2. Draw the full structure of two nucleotides - one from
DNA and one from RNA. Point out the key difference
between the two.
3. Write out an example of a condensation reaction forming
a phosphate-sugar bond.
4. Write out an example of a condensation reaction between
a nucleic acid base and a sugar.
5. State the "Central Dogma" of biochemistry.
arrow_forward
For the tripeptide ala-lys-phe
a) Draw the structure of the peptide you would expect at pH 1. Show all lone pairs of electrons and show formal charges. Determine the charge on the tripeptide.
b) Draw the structure of the peptide you would expect at pH 7. Show all lone pairs of electrons and show formal charges. Determine the charge on the tripeptide..
arrow_forward
Suppose that you were to construct an octapeptide utilizing seven valines and one
alanine. How many different octapeptides could you construct?
Question 10
Write the complementary DNA base sequence for the following DNA segment.
Indicate the 5' and 3' ends. (Type these in with no gaps - for example 5'BBZZ3')
5'GCATCATG3
arrow_forward
if the RNA molecule is 40 residues long and all four bases can be at any reidue, then the total number of RNA molecules 4^40. show complete calculations, including units
arrow_forward
01
g
indow
110
© Macmillan Learning
Help
achieve.macmillanlearning.com
3
X
Chapter 11 HW - General, Organic, and Biological C
Place an asterisks (*) next to the 3' carbon atoms in the polynucleotide shown.
O
O-P-O-CH2
Guanine
Answer Bank
H
H
Ο
H
O
H
Incorrect Answer
O P―0-CH2
Thymine
H
H
H
Cytosine
O
I-
O P―O-CH2
H
H
Ο
H
OH
H
What is the name of this polynucleotide?
O CGT
○ CTG
○ GTC
↓
TCG
NOV
21
Incorrect Answer
M
ப
A
Solution
Resources
Penalized
arrow_forward
How can we quantify the amount of energy required to generate a concentration gradient (Figure 13.1)?
arrow_forward
Kk.283.
arrow_forward
Select the single best answer.
Watson and Crick determined that the DNA molecule has two helical strands. Each strand is made up
which consist of a base, a deoxyribose, and a phosphate group linked together.
of
What word most correctly fills in the blank in the preceding statement?
nucleotides
fatty acids
monosaccharides
amino acids
arrow_forward
264&OpenVellumHMAC-ab644d9786730f4650f72638a7bd0c5b#10001
Part A
L
Submit
Part B
amylose and cellulose
Match the words in the left column to the appropriate blanks in the sentences on the right.
a(16)
B(14)
a(12)
glucose
a(14)
8(16)
a(12)
fructose
Request Answer
a(12)
B(16)
a(16)
cellulose and chitin
Match the words in the left column to the appropriate blanks in the sentences on the right.
glucose
-tose also called - Search
a(14)
fructose
8(14)
B(12)
x | +
Both are linear polysaccharides composed entirely of
linkages are
but in cellulose, they are
Both are linear polysaccharides with
2
DELL
and chitin contains N-acetylglucosamine.
A Q
Reset Help
In amylose, the glycosidic
glycosidic bonds. Cellulose contains
he
Reset Help
ENG
Constants
arrow_forward
Deoxyadenosine monophosphate (dAMP) and guanosine monophosphate (GMP) are nucleotides. The similarities between dAMP and GMP are that they both have?
-an alpha (central) carbon.-the same R group.-a phosphate group.-a pentose (5 sided) sugar-an amino group-a nitrogenous base.
arrow_forward
Youlube
Maps
EDD Debit Card - H...
Ahthem Blue Cross....
3rd Grade Mat...
Question 9.b of 20
A nucleotide contains a sugar, a phosphate, and a base. A picture of a nucleotide
is shown, deoxyadenosine 5'-monophosphate.
What type of functional group connects the phosphate to the sugar (shown in the
box)?
NH2
A) phosphodiester
B) ester
C) phosphoester
D) phosphoric acid
ОН
62°F
近
arrow_forward
Please don't provide handwritten solution ....
arrow_forward
3
arrow_forward
Question 31
Use the following chromatogram of standard amino acids:
5
24
11
4
16
19
3
12
15
23
6 8 9 14
13
14
17
18
20
22
1.
2
6 8
10
12
14
16
18
20 22
24
26
Time (min)
Which of the following choices describe structurally similar compounds in the chromatogram?
O Peak 11 and 24
O Peak 14 and 22
O Peak 12 and 21
O Peak 4 and 5
21
2.
2.
[nu)
arrow_forward
What is the final DNA concentration when a 10 μl of DNA at a concentration of 4
µg/μl is diluted to 50 μl with sddH₂O?
arrow_forward
This technique uses the principle of isoelectric point to separate a mixture of proteins into its pure constituents.Immersive Reader
electrophoresis
paper chromatography
X-ray diffraction
polymerase chain reaction
arrow_forward
Draw a complete lewis dot structure(showing all atoms,bonds,ione pairs, and charges) of the nucleotide DEOXYCYTIDYLATE and clearly circle the phosphoester bond in the structure. The base is Cytosine and the sugar is Deoxyribose.
arrow_forward
Protein sequence is always read (blank) terminal to (blank) terminal.
And DNA sequence is always read (blank) prime to ( blank) prime.
I need help filling in the blanks please? ? Thanks
arrow_forward
11. Draw the dinucleotide containing two molecules of 5'-monophosphate-guanosine in
the space below.
arrow_forward
Section one. Match enzyme or molecule with its function in bacteria. Answers may be used more than once or not at all.
DNA ligase
d. tRNA
promoter
а.
g.
b. DNA helicase
1RNA
h. terminator
е.
RNA polymerase
f.
DNA polymerase III
i.
stop codon
с.
37.
main enzyme in catalyzing DNA → mRNA transcription
38.
unwinds double-stranded DNA at the replication fork
39.
Where transcription stops
40.
adds new DNA bases in the 5'→3’ direction
41.
the "translator" molecule
42.
contains codons
43.
contains anti-codons
arrow_forward
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Related Questions
- questions 23-31arrow_forward#41 and #42arrow_forwardImportant steps in sequencing pure proteins include all of these except: a. Determining the amino acids on the ends of the smaller peptides b. Determining the amino acids on the ends of the protein c. Determining the isoionic pH of the protein d. Determining the amino acid composition e. Breaking the protein into smaller peptides It is impossible to sequence a protein if you do not have overlapping sequences to work with. A. True B. Falsearrow_forward
- Based on the following nucleotide structure, answer the following questions: H2N N- Но-Р-о- N. OH a. Is the base present in this nucleotide a purine or a pyrimidine? b. What is the abbreviation for the base present in this nucleotide? (A, T, C, G, U) Would the sugar in this nucleotide be found in molecules of DNA or RNA or both? d. Would the base in this nucleotide be found in molecules of DNA, RNA or both?arrow_forwardAre the following compounds correct? These may OR may not be correct. Please look them over and correct them if needed. Please list what was done incorrectly if applicable. b. Is the following nucleotide found in DNA? If not, please explain why. a. Is the following tRNA correct? NH2 VALINE <5' N. 3'- 0=P-0-CH2 ОН ОН GUA a. b. DNAA=T G=Carrow_forwardOn your Scantron, best possible answer for each question and fill in the corresponding bubble on your Scantron There are 50 total questions, each worth 3 points. 1. Which structure represents a nucleotide? A) CH,O- -O. 0- OH ОН B) NH2 CH, H H OH OH C) HO-CH, OH H H H. OH H H D) NH, 0. CH, OH OH 0-P=0 2. Which vitamin is fat soluble?arrow_forward
- nucleotides 2 Which component is not found in a nucleotide? Select an answer and submit. For keyboard navigation, use the up/down arrow keys to select an answer. phosphate b. base sulfate saccharidearrow_forwardThe DNA of bacteriophage lambda has 1.2 x 10^5 nucleotides. How many proteins of molecular weight 40,000 could be coded by this DNA? Assume a molecular weight of 100 for the average amino acid.arrow_forwardANALYSIS 1. A viral genome is composed of a double stranded DNA molecule containing 14% T (thymine). Based on this information, what would be the percentage of cytosine?arrow_forward
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