Homework Answers
Vector (300ng/uL)-------0.38ul(113.6ng)
Insert(600ng/uL)---------0.23uL(136.4ng)
Ligase mix(400U/uL)--1.0Um
Water--------------------------3.39uL
Total-----------------5uL
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3. The DNA ligase comes pre-mixed in its buffer in a 2x fold concentration, and your...
Your lab instructor has given you a protocol to perform a molecular cloning experiment. In a...
Your lab instructor has given you a protocol to perform a molecular cloning experiment. In a previous experiment, you used polymerase chain reaction (PCR) to amplify a sequence that you believe to regulate expression of a gene you are studying. You will now take this purified PCR product (double stranded DNA) and ligate it into a plasmid that contains a luciferase reporter gene. If your DNA sequence is a promoter sequence, then its presence will allow for expression of the...
Question 5 Using stock solutions in a protocol: from volume to final concentration A DNA ligation...
Question 5 Using stock solutions in a protocol: from volume to final concentration A DNA ligation reaction is carried out in a 25 μL reaction mix. This is a reaction carried out in cloning. You are going to do a number of these reactions so you decide to make up a larger volume of the reaction mixture, called a master mix. This saves on multiple repetitive pipetting and reduces errors. You have been given a protocol to make up 1...
You extracted your DNA from cheek cells. The concentration of your DNA sample is 250...
You extracted your DNA from cheek cells. The concentration of your DNA sample is 250 ng/?l. The DNA template for PCR should be 10 ng/?l. You should mix _____ ?l of your DNA sample and _____ ?l of water to make final volume of 100 ?l of DNA that has the concentration of 10 ng/?l. ?
4 total reactions 10. Complete this Master Mix table for 3 DNA samples, a positive control,...
4 total reactions
10. Complete this Master Mix table for 3 DNA samples, a positive control, negative control, and an extra reaction for pipetting error. Show your work. 4 pts) Master C Mix Conc. Of Final Stock solution L/Rxn Number of Total L (total per Reactions needed tube) for master mix PCR buffer Water dNTP mix MgCl2 F Primer 20X 1X | | 25 mM2.5 mM 10uM | 0.1 μΜ 100 ml 100 μΜ RPrimer | 10uM | 0.1μΜ Taq...
Question 7 Working with buffers You need 100 ml of 120 mM phosphate buffer and you have two stock...
please hlep me answer those three questions asap.
Please ignore question 7.
Question 7 Working with buffers You need 100 ml of 120 mM phosphate buffer and you have two stock solutions from which you can make the buffer; 0.6 M NaH2PO4 (the acid form) and 0.6 M Na2HP04 (the base form). You need to add 3 times more of the base form than the acid form to achieve the desired pH. How will you make this solution? Question 8...
See image please show full solution, (Indicate your answers to two digits after the decimal) 8....
See image
please show full solution,
(Indicate your answers to two
digits after the decimal)
8. You have two plasmid preparations "A and "B" (illustrated below) each at concentrations of 100nguL. You wish to subclone a 1.5 Kbp EcoRI fragment from plasmid·A" into the unique EcoRI site of a 3.0Kbp vector"B". EcoRI 1.5 Kbp Total size 3.0Kbp BT Total size 7.5Kbp Complete the following table to indicate the volumes of each ingredient required for the preparation of a 50μL ligation...
You are preparing to clone a DNA fragment into a plasmid vector. You start by linearizing...
You are preparing to clone a DNA fragment into a plasmid vector. You start by linearizing your plasmid (concentration = 500 ng/μl) with EcoRI, which is provided in a standard 50% glycerol solution at 10 units/μl. Your enzyme also comes with an appropriate 10X reaction buffer. Taking into account the final allowable glycerol concentration, you want to add the maximum amount of EcoRI, to achieve complete digestion of 1 μg plasmid DNA in a total volume of 20 μl. Fill...
2. Calculate the final concentration of yeast and buffer in each of the four reaction tubes....
2. Calculate the final concentration of yeast and buffer in each of the four reaction tubes. Enter your results in Table 1 below. Table 1. Standard Fermentation Reactions 1 pt Reagents Reaction mix (ml) Tube #1 Tube #2 Tube #3 Tube #4 Water 7.0 6.0 4.0 5.0 0.2 M NaP buffer, pH 7 2.0 2.0 2.0 2.0 60% glucose 1.0 1.0 1.0 0 __% yeast 1.0 3.0 3.0 Total volume of each reaction 10.0 10.0 10.0 Final concentration of glucose...
I just need the answers to questions 2 and 3. My DNA ladder is in lane...
I just need the answers to questions 2 and 3. My DNA ladder is
in lane 2 with the yellow arrow pointing to it. Thanks!
Part 2: Gel purification and ration Gel Slice and PCR Product Preparation modified from IBSci.com instructions for gel and PCR clean-up system A. Dissolving the Gel Slice 1. Following electrophoresis, excise DNA band from gel and place gel slice in a 1.5ml microcentrifuge tube. 1b. Use an analytical balance to weigh gel slice. Record weight...
I need the answers for questions 2 and 3. My DNA ladder is in lane 2...
I
need the answers for questions 2 and 3. My DNA ladder is in lane 2
marked by the yellow arrow. Thanks!
Here is the only other info I have. Thanks!
Part 2: Gel purification and on Gel Slice and PCR Product Preparin modified from TBSci.com instructions for gaan A. Dissolving the Gel Stie Following electrophores, eral DNA band from grand place glice microcentrifuge tube Ib. Use an analytical balance to weigh pelice Rec die 2. Add 500 balance to...
Question 5 Using stock solutions in a protocol: from volume to final concentration A DNA ligation reaction is carried out in a 25 μL reaction mix. This is a reaction carried out in cloning. You are going to do a number of these reactions so you decide to make up a larger volume of the reaction mixture, called a master mix. This saves on multiple repetitive pipetting and reduces errors. You have been given a protocol to make up 1...
4 total reactions
10. Complete this Master Mix table for 3 DNA samples, a positive control, negative control, and an extra reaction for pipetting error. Show your work. 4 pts) Master C Mix Conc. Of Final Stock solution L/Rxn Number of Total L (total per Reactions needed tube) for master mix PCR buffer Water dNTP mix MgCl2 F Primer 20X 1X | | 25 mM2.5 mM 10uM | 0.1 μΜ 100 ml 100 μΜ RPrimer | 10uM | 0.1μΜ Taq...
please hlep me answer those three questions asap.
Please ignore question 7.
Question 7 Working with buffers You need 100 ml of 120 mM phosphate buffer and you have two stock solutions from which you can make the buffer; 0.6 M NaH2PO4 (the acid form) and 0.6 M Na2HP04 (the base form). You need to add 3 times more of the base form than the acid form to achieve the desired pH. How will you make this solution? Question 8...
See image
please show full solution,
(Indicate your answers to two
digits after the decimal)
8. You have two plasmid preparations "A and "B" (illustrated below) each at concentrations of 100nguL. You wish to subclone a 1.5 Kbp EcoRI fragment from plasmid·A" into the unique EcoRI site of a 3.0Kbp vector"B". EcoRI 1.5 Kbp Total size 3.0Kbp BT Total size 7.5Kbp Complete the following table to indicate the volumes of each ingredient required for the preparation of a 50μL ligation...
2. Calculate the final concentration of yeast and buffer in each of the four reaction tubes. Enter your results in Table 1 below. Table 1. Standard Fermentation Reactions 1 pt Reagents Reaction mix (ml) Tube #1 Tube #2 Tube #3 Tube #4 Water 7.0 6.0 4.0 5.0 0.2 M NaP buffer, pH 7 2.0 2.0 2.0 2.0 60% glucose 1.0 1.0 1.0 0 __% yeast 1.0 3.0 3.0 Total volume of each reaction 10.0 10.0 10.0 Final concentration of glucose...
I just need the answers to questions 2 and 3. My DNA ladder is
in lane 2 with the yellow arrow pointing to it. Thanks!
Part 2: Gel purification and ration Gel Slice and PCR Product Preparation modified from IBSci.com instructions for gel and PCR clean-up system A. Dissolving the Gel Slice 1. Following electrophoresis, excise DNA band from gel and place gel slice in a 1.5ml microcentrifuge tube. 1b. Use an analytical balance to weigh gel slice. Record weight...
I
need the answers for questions 2 and 3. My DNA ladder is in lane 2
marked by the yellow arrow. Thanks!
Here is the only other info I have. Thanks!
Part 2: Gel purification and on Gel Slice and PCR Product Preparin modified from TBSci.com instructions for gaan A. Dissolving the Gel Stie Following electrophores, eral DNA band from grand place glice microcentrifuge tube Ib. Use an analytical balance to weigh pelice Rec die 2. Add 500 balance to...
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