Docsity
Docsity
Prepare for your exams
Prepare for your exams

Study with the several resources on Docsity

Earn points to download
Earn points to download

Earn points by helping other students or get them with a premium plan

Guidelines and tips
Guidelines and tips
Sell on Docsity
Sell on Docsity
Docsity AI
Docsity AI
Log inSign up

Prepare for your exams

Study with the several resources on Docsity

Find documents

Prepare for your exams with the study notes shared by other students like you on Docsity

Search for your university

Find the specific documents for your university's exams

Docsity AINEW

Summarize your documents, ask them questions, convert them into quizzes and concept maps

Explore questions

Clear up your doubts by reading the answers to questions asked by your fellow students

Earn points to download

Earn points by helping other students or get them with a premium plan

Share documents
download point icon20 Points

For each uploaded document

Answer questions
download point icon5 Points

For each given answer (max 1 per day)

All the ways to get free points
Get points immediately

Choose a premium plan with all the points you need

Study Opportunities

Choose your next study program

Get in touch with the best universities in the world. Search through thousands of universities and official partners

Community

Ask the community

Ask the community for help and clear up your study doubts

Free resources

Our save-the-student-ebooks!

Download our free guides on studying techniques, anxiety management strategies, and thesis advice from Docsity tutors

Homework chapters 13-18, Assignments of Quantitative Techniques

University of Central Florida (UCF)Quantitative Techniques

Homework chapters 13-18 2022 fall

Typology: Assignments

2021/2022

Uploaded on 02/04/2023

mohammad-jamous
mohammad-jamous 🇺🇸

1 document

1 / 6

Toggle sidebar

This page cannot be seen from the preview

Don't miss anything!

bg1
Chapters 13-18 Homework Name: _______________________________________ Lab Section: _______ Seat: _______
Show work, and enter final answer on the line provided where applicable.
Chapter 13:
1. The following fragments of DNA are run
on a gel. Label the marker (M) and write
the fragment size at the top of each lane.
DNA Fragments:
1800 bp 600 bp
1100 bp 400 bp
900 bp 100 bp
2. You digest a 1000 bp plasmid with HindIII, and run a linear marker and 0-20 minute time points of this digestion on a
gel. Which type of DNA is found in each row? Write the answer on the arrows to the right of the gel.
Use the following terms:
Supercoiled Circular
Linear Nicked
Chapter 14:
3. A restriction map of a linear 1 kb piece of DNA is shown. Draw the bands you would expect to see on the gel if the
DNA was completely digested with the restriction enzymes indicated at the top of each lane.
a.
Mohammad Jamous
0020
9A
M
600
100
1800
400
1100
900
Circular
Nicked
Linear
Supercoiled
800
600
500. 500
400. 400. 400
300. 300. 300
200. 200. 200
100. 100. 100. 100
pf3
pf4
pf5

Partial preview of the text

Download Homework chapters 13-18 and more Assignments Quantitative Techniques in PDF only on Docsity!

Chapters 13 - 18 Homework Name: _______________________________________ Lab Section: _______ Seat: _______ Show work, and enter final answer on the line provided where applicable. Chapter 13:

  1. The following fragments of DNA are run on a gel. Label the marker (M) and write the fragment size at the top of each lane. DNA Fragments: 1800 bp 600 bp 1100 bp 400 bp 900 bp 100 bp
  2. You digest a 1000 bp plasmid with HindIII, and run a linear marker and 0-20 minute time points of this digestion on a gel. Which type of DNA is found in each row? Write the answer on the arrows to the right of the gel. Use the following terms: Supercoiled Circular Linear Nicked Chapter 14:
  3. A restriction map of a linear 1 kb piece of DNA is shown. Draw the bands you would expect to see on the gel if the DNA was completely digested with the restriction enzymes indicated at the top of each lane. a.
  1. A 1 kb linear piece of DNA is digested with PstI, HpaI, and XbaI. Based on the gel, circle the letters next to the restriction maps on the right if they represent the locations of these digestion sites. Circle all that apply (more than one is correct).
  2. A 1 kb linear piece of DNA is digested with EcoRI and BamHI. Based on the gel, draw the location of these enzymes to create a restriction map (write “E” or “B” above the line at the digestion sites).
  3. A restriction map of a 1 kb circular vector is shown. Draw the bands you would expect to see on the gel if the DNA was completely digested with the restriction enzymes indicated at the top of each lane.
  4. You purify DNA from a miniprep and wish to digest it with EcoRI. You have DNA with an A 260 of 2.0, EcoRI at 1 0, units/ml (1 unit = 1 g of DNA digested per hour under optimal conditions, but 10 units are usually used per reaction to ensure complete digestion), 5X reaction buffer, and 10X BSA. Calculate the concentration (a) and fill in the table (b-e) to calculate how to prepare a restriction enzyme digestion of 1 g of DNA in 50 l. a. Using the A 260 , what is the DNA’s concentration? ____________ g/ml b. Using (a), how many l of DNA would equal 1 g? Enter (b-e) in the table. c. How many l of 5X reaction buffer should be added to the 50 l reaction? d. How many l of 10X BSA should be added to the 50 l reaction? e. 10 units of EcoRI (10,000 units/ml) must be used. How many l is this? Reagent (^) Volume (l) (b) 1 g DNA (c) 5X Buffer (d) 10X BSA (e) 10 units EcoRI Water Total: (^50) l
  1. A gene is inserted into the plasmid below using the following restriction enzymes. Circle the expected results (blue, white, or no growth), and explain your answer. Assume the plate contains ampicillin, IPTG, and X-gal. a. EcoRI and BamHI B W NG ___________________________________________ b. NcoI and NdeI B W NG ___________________________________________ c. XbaI and NdeI B W NG ___________________________________________ d. BamHI and PstI B W NG ___________________________________________ e. PstI and XbaI B W NG ___________________________________________ f. BamHI and HindIII B W NG ___________________________________________ g. Which enzymes ensure insert directionality and white cells? ____________________ Chapter 16:
  2. A family with a history of a genetic disorder were analyzed with a dot blot using a probe for both the normal and mutant allele. Color in the circles below each person to indicate the types of spots you would see on this dot blot.
  3. Draw the expected RFLP results for the three individuals shown (1-3) on the gel. Enter the genotypes of each person below the gel ( e.g. +/+) if the internal cut site correlates to a dominant genotype (+). a. _____ _____ _____
  4. A 100 bp VNTR exists at a locus in the human genome. Two parents have four children with different genotypes at this locus. Draw the bands you would expect to see on a gel for both the parents and all four children, and the genotype below each ( e.g. 3/4). a. _____ _____ _____ _____ _____ _____ b. The parents have a fifth child, with genotype 5/5. What are some possible explanations for this?
  1. Huntington’s Disease, an autosomal dominant disorder, was linked to a HindIII digestion site using a Southern Blot and a specific probe. Researchers detected four different digestion patterns, three of which are below (called haplotypes A, B, and C). Haplotype A is linked to the defective huntingtin gene, which causes neurodegeneration. a. Which individuals (#1-7) have Huntington’s Disease? _____________ b. Which band size on the gel represents haplotype B? _____________ c. A person has bands only at 3.7/1.2 kb (not 4.9 kb) but they don’t have the disease. What is their genotype ( e.g. A/A)? _____________ Chapter 17:
  2. Design forward and reverse primers to the following DNA fragments. Primers are typically ~20 nucleotides in length with annealing temperatures ~50-60 °C, but only design these primers to be six nucleotides for simplicity, and write all primers 5’→3’. Also calculate each primer’s Tm and the annealing temperature that should be used in PCR. a. 5’-GTACTAGTAGACCGTATGCCGCAGTA-3’ Forward: 5’-_____________________________ Tm: _______ Reverse: 5’-_____________________________ Tm: _______ Annealing Temperature: _______ b. 5’-GCTGTAAGAGGATTTACTACCGTCCA-3’ Forward: 5’-_____________________________ Tm: _______ Reverse: 5’-_____________________________ Tm: _______ Annealing Temperature: _______
  3. You wish to amplify the DNA fragments below and add cut sites for subsequent cloning. Design both primers with the restriction enzymes cut sites listed, and add two overhang bases to each primer. Label the extra bases and cut sites. Each primer should be 14 nucleotides in length. a. 5’-CTATGAGGTCCTGCGTTAGTGTTACC-3’ Forward: 5’-_____________________________ Tm: _______ 5’ EcoRI, 3’ BamHI, overhang bases Reverse: 5’-_____________________________ Tm: _______ Annealing Temperature: _______ b. 5’- CCTGGGTGTTACATGCCATTAGCGTT-3’ Forward: 5’-_____________________________ Tm: _______ 5’ HindIII, 3’ SphI, overhang bases Reverse: 5’-_____________________________ Tm: _______ Annealing Temperature: _______
  4. Design the internal forward and reverse primers to mutate the amino acids shown. Primers are typically ~ nucleotides in length, but only design these primers to be nine nucleotides. Label the mutated codon. a. 5’-TAAGGCTAGTAG ACC GTATGCCGAATAC-3’ Internal Forward: 5’-______________________ Tm: _______ Mutate the threonine to cysteine (TGC) Internal Reverse: 5’-______________________ Tm: _______ Annealing Temperature: _______ b. 5’-CGCGTATGAGAG GAT CTACTACGGATAG-3’ Internal Forward: 5’-______________________ Tm: _______ Mutate the aspartate to tyrosine (TAT) Internal Reverse: 5’-______________________ Tm: _______ Annealing Temperature: _______