Homework Answers
1. Forward primer: 5'-GCC-3'
Forward primer sequence will be same as that of the given sequence
and it binds to the complementary strand.
Reverse primer: 5'-GAT-3'
reverse primer sequence will be complementary to that of the given
sequence and it binds to the given strand.
2. A single band indicates the presence of a single allele i.e.
homozygosity.
Two bands indicate the presence of two different alleles i.e.
heterozygosity.
Patient 1= Homozygous
Patient 2= Heterozygous
Patient 3= Homozygous
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based on the PCR results, estimate how mang CTG repeats are
present in rhe DMPK gene...
1. Create the primers to amplify the gene of interest. 1. Below is almost the full...
1. Create the primers to amplify the gene of interest. 1. Below is almost the full sequence of the coding strand of the F9 gene (exon 2-exon4) that you found in the online database (NCBI, Genomic sequence F9 gene). Some of the sequence is missing, but you know how many nucleotides are skipped. Design forward and reverse primers 18nt each for PCR that will isolate EXACTLY the sequence that is in bold typeface. Exon 2 = 164 nt Intron 2...
2. PCR amplification of the TAS2R38 gene a. The number of copies of the 303 bp sequence grows exp...
2. PCR amplification of the TAS2R38 gene a. The number of copies of the 303 bp sequence grows exponentially (1-2-4-8-etc) after each cycle. The number of cycles we used is on page 97. What is the number of copies of the 303 bp fragment that will theoretically be present at the end of our reaction? b. Denaturation of the 303 bp segment of the TAS2R38 gene is a critical first step in the PCR perties of a DNA segment that...
You are using PCR to amplify a 300 bp target sequence, a portion of Gene X,...
You are using PCR to amplify a 300 bp target sequence, a portion of Gene X, from human genomic DNA isolated from patients' blood samples. The instructions for this procedure tell you to include Samples A and B, whose contents are listed below, with each batch of patient samples that you run. Ingredients Sample A Sample B 10x PCR Buffer (Tris,KCI,MgCl2,BSA) 5 mL 5 mL H2O 37.8mL 38.8mL dNTP's 3 mL 3 mL Taq DNA polymerase 0.2 mL 0.2 mL...
based on the given graph how would these be answered? sa sequence from a sheep PCR...
based on the given graph how would these be answered?
sa sequence from a sheep PCR product. The template used was genomic DNA and the forward and se primers were designed to amplify partofthe priongene, which encodes the causative agentot scrapie. is black;Cisblue; Ais green: Tisred. The forward primerwasusedasasequencing primer. TheNattheten ow means that the computer can't decide which nucleotide is at that position in the sequence because there are two peaks (red=T and blue-C) that are the same height....
Carolina Savirana Craz 3/12/20 GECC-Polymerase Chain Reaction 1. What is the purpose of the polymerase chain...
Carolina Savirana Craz 3/12/20 GECC-Polymerase Chain Reaction 1. What is the purpose of the polymerase chain reaction? a. To repair damaged DNA b. To make copies of entire chromosomes c. To make copies of specific regions of DNA d. To prepare cells for cell division 2. The polymerase chain reaction is most comparable to what cellular process? a. Mitosis b. Replication c. Transcription d. Translation 3. When enzymes are elongating (building) a newly synthesized DNA strand in PCR, new nucleotides...
QUESTION 1: You are inserting a gene into an MCS found within the LacZ gene. Using...
QUESTION 1: You are inserting a gene into an MCS found within the LacZ gene. Using blue/white colony selection, why could you assume that white colonies have modified plasmids? a. A blue colony means the LacZ reading-frame was disrupted b. A blue colony means your gene has mutations c. A white colony means the LacZ reading-frame is intact d. A white colony means the LacZ reading-frame was disrupted QUESTION 2: You are performing a PCR using primers with a sequence perfectly...
We understand that your diagram will not be able to portray the bands to their exact base-pair si...
We understand that your diagram will not be able to portray the bands to their exact base-pair size, and that in some cases you may not be able to calculate an exact size for your bands. Just do your best in terms of positioning the bands in the lanes with respect to the standard markers, but please do not panic if the bands are a couple of mm away from perfect in your diagram. LABEL each band with its size...
Background: Human Epidermal Growth Factor (h-EGF) is produced in small quantities by glands all over the...
Background: Human Epidermal Growth Factor (h-EGF) is produced in small quantities by glands all over the body and transported to the epidermis. Epidermis cells undergo a rapid turnover and new cells are constantly needed to replace the old epidermis cells as they die. h-EGF facilitates the constant low-level production of epidermis cells. While most people make sufficient quantities of h-EGF for their own skin regenerations, patients with large wounds or burns benefit from a treatment using h-EGF to stimulate epidermis...
Based on the results from the agarose PCR (240-bp product), how many alanines are likely to...
Based on the results from the agarose PCR (240-bp product), how
many alanines are likely to be inserted in the PHOXB
sequence?
a. 30–40
b. 5–8
c. 15–20
d.
8–12
Case Study 12-2 A 7-month-old child suffered from sleep apnea and difficulty breathing. Respiratory infections and allergies were ruled out. From the family history, the physician suspected congenital congestive hypoventilation syndrome (CCHS). CCHS results, in part, from abnormal function of the PHOX2B gene product caused by a triplet-repeat expansion. The...
8. Design appropriate PCR primers for the human DDX3X gene. Give their sequences below and explain...
8. Design appropriate PCR primers for the human DDX3X gene. Give their sequences below and explain how you designed them (20 points, 10 each). The DDX3X cDNA sequence from the NCBI database is provided to you on Moodle Within the sequence, first identify the coding sequence (cds) as this is the region you want to amplify by PCR Hint: It tells you on the first page of the sequence file where the cds is positioned! Also remember that a cds...
1. Create the primers to amplify the gene of interest. 1. Below is almost the full sequence of the coding strand of the F9 gene (exon 2-exon4) that you found in the online database (NCBI, Genomic sequence F9 gene). Some of the sequence is missing, but you know how many nucleotides are skipped. Design forward and reverse primers 18nt each for PCR that will isolate EXACTLY the sequence that is in bold typeface. Exon 2 = 164 nt Intron 2...
2. PCR amplification of the TAS2R38 gene a. The number of copies of the 303 bp sequence grows exponentially (1-2-4-8-etc) after each cycle. The number of cycles we used is on page 97. What is the number of copies of the 303 bp fragment that will theoretically be present at the end of our reaction? b. Denaturation of the 303 bp segment of the TAS2R38 gene is a critical first step in the PCR perties of a DNA segment that...
based on the given graph how would these be answered?
sa sequence from a sheep PCR product. The template used was genomic DNA and the forward and se primers were designed to amplify partofthe priongene, which encodes the causative agentot scrapie. is black;Cisblue; Ais green: Tisred. The forward primerwasusedasasequencing primer. TheNattheten ow means that the computer can't decide which nucleotide is at that position in the sequence because there are two peaks (red=T and blue-C) that are the same height....
Carolina Savirana Craz 3/12/20 GECC-Polymerase Chain Reaction 1. What is the purpose of the polymerase chain reaction? a. To repair damaged DNA b. To make copies of entire chromosomes c. To make copies of specific regions of DNA d. To prepare cells for cell division 2. The polymerase chain reaction is most comparable to what cellular process? a. Mitosis b. Replication c. Transcription d. Translation 3. When enzymes are elongating (building) a newly synthesized DNA strand in PCR, new nucleotides...
We understand that your diagram will not be able to portray the bands to their exact base-pair size, and that in some cases you may not be able to calculate an exact size for your bands. Just do your best in terms of positioning the bands in the lanes with respect to the standard markers, but please do not panic if the bands are a couple of mm away from perfect in your diagram. LABEL each band with its size...
Based on the results from the agarose PCR (240-bp product), how
many alanines are likely to be inserted in the PHOXB
sequence?
a. 30–40
b. 5–8
c. 15–20
d.
8–12
Case Study 12-2 A 7-month-old child suffered from sleep apnea and difficulty breathing. Respiratory infections and allergies were ruled out. From the family history, the physician suspected congenital congestive hypoventilation syndrome (CCHS). CCHS results, in part, from abnormal function of the PHOX2B gene product caused by a triplet-repeat expansion. The...
8. Design appropriate PCR primers for the human DDX3X gene. Give their sequences below and explain how you designed them (20 points, 10 each). The DDX3X cDNA sequence from the NCBI database is provided to you on Moodle Within the sequence, first identify the coding sequence (cds) as this is the region you want to amplify by PCR Hint: It tells you on the first page of the sequence file where the cds is positioned! Also remember that a cds...
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