What is the correct order to prepare pure recombinant proteins when using E coli expression system? 1. Select appropriate column chromatography to purify the recombinant proteins
Q: You observe the following: Your Your friend's Gel Gel 120 kDa 100 kDa 120 kDa 100 kDa -80 kDa 80 kDa…
A: you have asked multiple question please repost the question and mention the sub parts needed to be…
Q: You observe the following: Your Your friend's Gel Gel 120 kDa 100 kDa 120 kDa -100 kDa -80 kDa 80…
A: Blotting, as we all know, is a process that uses electrophoresis to separate DNA, RNA, and protein…
Q: Below is a diagram of the vector you are planning to use. You identify four restriction enzyme…
A: Restriction enzymes are the cleaving enzymes and thus are also known as molecular scissors. These…
Q: A thermophillic bacterium was isolated from a hot spring at Perak to study its thermostable amylase…
A: DNA technology is the chemical manipulation of the genotype and the resulting phenotype of organisms…
Q: Discuss how scientists employ a bacteriophagesite-specific recombination system to generateknockin…
A: Knockin mouse is a method in gene recombination in which an exogenous gene of interest is inserted…
Q: 1. Restriction enzymes are extensively used in molecular biology. Below are the recognition sites of…
A: Overhangs are single stranded ends of the DNA nucleotides which are formed by when the DNA is…
Q: Which of the following statements about base editing is correct? 1. It involves nCas9. 2. The end…
A: Base editing It is an alternative tool to HDR-mediated replacement. This facilitates editing of…
Q: How do you do a DNase footprint assay. What did it demonstrate in regards to the UP elements binding…
A: The DNase footprinting technique is a DNA footprinting technique. It cannot be confused with…
Q: Primer design for SLIC cloning and the protocol
A: Hi! Thank you for the questions. As you have posted multiple questions, I will be answering the…
Q: To quantify the transcription product of a specific gene in E. coli, you may use (select all correct…
A: Introduction The Process Of Transcribing A Piece Of DNA Into RNA Is Known As Transcription.…
Q: 4. The image below is based on the Meselson Stahl experiment where bacteria are grown in 15N and…
A: The Meselson-Stahl experiment showed that DNA replication is a semiconservative process. In this…
Q: Consider a bacterial promoter with -35 and -10 elements. What assay is best to show that RNA…
A: The electrophoretic mobility shift assay (EMSA) is a method for the detection of protein-nucleic…
Q: Amplified target regions of four different samples were separated using gel electrophoresis. DNA…
A: Gel electrophoresis is a lab technique for separating DNA, RNA, and protein mixture based on their…
Q: following site does the restriction enzymes act? 32. Restriction Fragment Length Polymorphism (RFLP)…
A: Restriction endonucleases are enzymes that cuts the DNA at specific DNA sequence known as…
Q: Coding With the given coding strand perform the following 1. supply the correct non- coding strand…
A: DNA is a double helical molecule.
Q: Once you have idenified a gene that you suspect is responsible for the disease, you clone the gene…
A: Out of all the options given - Option - Northern Blot , is incorrect because northern blotting is a…
Q: B. Arrange the following steps in the genetic engineering of corn to acquire pest resistance. to…
A: Genetic engineering is modification in the genetic makeups of the organism in order to get good…
Q: 9) Describe the steps required to use sequencing DNA to identify your gene of interest after…
A: DNA sequencing is the process of finding the sequence of the nucleotides on a particular short…
Q: In organism that is mutant for components of the non-homologous end-joining NHEJ repair pathway…
A: CRISPR gene editing is a molecular biology genetic engineering tool for altering the genomes of…
Q: These are the results after running Agarose Gel Electrophoresis of the cut/uncut pUC19 plasmid and…
A: These are the results after running Agarose Gel Electrophoresis of the cut/uncut pUC19 plasmid and…
Q: How can the efficiency of homologous recombination be improved when using CRISPR-Cas9, compared to…
A: CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is a technique derived from…
Q: robes for cloned genes use ____. a. certain bacteria that glow when they take up the genes b.…
A: Gene cloning: It is a procedure that makes researchers prepare several similar copies of gene-sized…
Q: 1. An 9 kb circular plasmid is cut with the EcoRI restriction enzyme, and the reaction products an…
A: An inclusion could be a tiny, circular, double-stranded DNA molecule that's distinct from a cell's…
Q: Explain how site directed mutagenesis can be accomplished using M13 bacteriophage. Using this method…
A: Deoxyribonucleic acid (DNA) is a hereditary molecule that passes genetic information from one…
Q: 2. You are planning to clone a gene into the PBR322 plasmid vector. The gene has Bgl II restriction…
A: Gene cloning is the technique to produce the exact copies of the desired gene by utilizing genetic…
Q: The human gene for hemophilia is on the X chromosome. Below is a pedigree from a family afflicted…
A: Hi, Thanks For Your Question. Answer : Correct Option Is b (20%) Explanation : P (III-7 Is…
Q: The DNA sequence of one strand of a gene from threeindependently isolated mutants is given here (5′…
A: The mutant gene produces when there are changes in the sequence of nucleotides in the wild type…
Q: To quantify the transcription product of a specific gene in E. coli, you may use (select all correct…
A: Transcription It is a biological processe where RNA is made by using DNA as a template.
Q: The concentration of your RNA solution is 1500 ng/μl. How much RNA solution do you need to use when…
A: The concentration of RNA in solution can be determined by measuring absorbance at 360 nm.
Q: Select the best technique to answer the following question: Where does the transcription factor…
A: The Nanog protein activator for the Rex-1 promoter by positive feedback mechanism and it's playing a…
Q: 2. can you conceive of any potential applications of CRISPR technology?
A: can you conceive of any potential applications of CRISPR technology? Ans -CRISPR (also referred to…
Q: Sequencing reactions are done in separate tubes for each ddNTP with a radioactive primer. Which…
A: Gell electrophoresis is a biological apperatus mainly used in biotechnology or in case of genetic…
Q: The method that Mandla should use to distinguish recombinants from non-recombinants is called th…
A: Recombinant DNA (rDNA) molecules are DNA molecules created by laboratory genetic recombination…
Q: 7:12 What is not true for Sequence tagged site (STS) markers: O cannot be mapped by fluorescence in…
A: Any sequence of DNA which shows polymorphism and can be recognized utilizing a molecular method is…
Q: During conjugation of an F strain of E. coli with an Hfr strain, it takes about 100 minutes to…
A: Bacterial conjugation is the process of transfer of genetic material between two cells by a…
Q: In bacterial transformation ... 1, Why do you have to incubate the competent bacterial cells on ice…
A: The process which aids the bacteria in taking up a specific naked DNA (deoxyribonucleic acid) from…
Q: Describe how to select recombinant clones if a foreign DNA is inserted into the polylinker site of…
A: Introuction- pUC18 is a 2686 bps long plasmid vector which contains pMB1 as origin of replication…
Q: Choose the proper order for recombinant protein purification from bacterial cells. a.…
A: Recombinant proteins are a new combination of genes that forms DNA. It allows for the production of…
Q: Explain the process of how X-gal screening works with pUC19, you may build a model with boxes and…
A: Hello. Since your question has multiple parts, we will solve the first question for you. If you want…
Q: 1. The gel presented here shows the pattern of bands of fragments produced with several restriction…
A: The various fragments obtained in the gel are: Aatl 1 → 9kb, 3kb = 12kb.Ncol → 8kb, 4kb.Aatl 1 +…
Q: What size bands would you expect to see on the Southern blots using probe 1 and probe 2 for an…
A: Introduction The term Sickle cell disease defines the condition where due to a mutation, the blood…
Q: Explain your experimental approach if you must correct the genetic defect phenylketonuria (PKU),…
A: Phenylketonuria is a genetic defect that is caused by the deficiency of phenylalanine hydroxylase…
Q: Ligation is an essential step in the cloning process. It refers to the joining of the gene of…
A: According to the question, we have to determine the four control groups which are important to…
Q: SDS-PAGE is used to help determine all of the following except: A) The purity of the recombinant…
A: A polyacrylamide gel consists of chains of acrylamide monomers cross-linked with N,…
Q: You inserted your gene of interest in pBR322 as illustrated below. You spliced the vector and insert…
A: Screening The procedure to identify and select the individual of interest.
Q: Draw a gel to represent the band shift assay result for testing the following mixtures: (1)…
A: Band shift assay is also known as electrophoretic mobility shift assay (EMSA) is the technique used…
Trending now
This is a popular solution!
Step by step
Solved in 2 steps
- A cloning vector map is shown below. EcoRI Bam Ban Hind P-galactosidase Amp Bam Bam EcoRI Ori C Which restriction site is best for inserting a DNA fragment for selection of chimeric plasmid containing colonies? 1) They're all equally good. 2) Hindll 3) EcoRI 4) BamHIKha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHIDuring which step in the transformation protocol does the plasmid DNA move into the cell? Heat Shock Grow in LB media at 37°C Plate on media with Ampicillin O Incubate the cells on ice with CaCl2 in the absence of plasmid. fg f1o 19 %24 & 5 6. 7 8. 4 5 R Y U P F G J 2 K 3] vの
- 5 μL of plasmid DNA (pUC 19) was added to 50ul of CaCl2 competent E. coli that was then added 250 μL of SOC medium, 100 μL of this solution was plated onto a TSA + ampicillin plate. Can you please show me how I calculate the total number of successful transformants per mL of competent cells plated. The total number of colnies counted on plate was 70. Thank youOne of the frist steps in isolating plasmid DNA via mini-prep is to pellet the cells after O/N culture and then resuspend them in buffer P1. What's the point of pelleting the cells just to resuspend them again? To concentrate the cells P1 is a buffer, preventing small changes in pH O This step is not absolutely necessary P1 begins the lysis process D Question 2 What would happen if we didn't centrifuge the tube containing lysed cells after adding neutralization buffer and instead just added right to the column? O We could fihish the prep but we would have protein/RNA contaminants at the end O The precipitate is less dense than water so it shouldn't get in the way since it floats on top O The precipitate would obstruct the column and our prep is ruined O Our yield would be reduced, but we'd get somethingHelp Please Supercoiled DNA migrates through an agorose gel faster than similarly sized linear DNA? True or False? Using the plasmid map, indicate where (bp region on plasmid) these enzymes cut on the plasmid. EcoR1 1. 1 Hindlll 2. 637 Bpu101 3. 14 BamHI 4. 256 To sequence a DNA plasmid, you need 1 µg of DNA. If your DNA concentration was 0.45 µg/µL. How many µl of your DNA 0.45 µg/µL is required to get 1 µg final? Answer in µl using 2 decimal places. Your Answer: Answer units >
- 3A. Following transformation with the CRISPR plasmid, the yeast will be plated on SD-URA plates. Why is SD-URA media used? Name the relevant marker gene in the pCRCT CRISPR plasmid. 3B. What would you expect to see if you plated on YED accidentally? Will most of the yeast be red or white? Why? 3C. The complete genotype of the host yeast strain is MATa ade2 his3 leu2 met15 ura3. Name one advantage of using a yeast strain with auxotrophies for several genes (i.e. his3, leu2 and ura3). 3D. After successful CRIPSR editing, the yeast can later be “cured” of the recombinant CRISPR plasmid – that is, the plasmid is lost but the CRISPR edit is stably inherited. Write the new genotype of the yeast-based on the ADE6 gene.Competent E. coli cells were transformed with the pGLO plasmid. These transformed cells were then allowed to grow on two different plates: 1) a plate containing LB/AMP and 2) another plate containing LB/AMP/ARA. In which plate would you observe both phenotypic and genotypic changes? Briefly justify your answer. Edit View Insert Format Tools Table 12pt v Paragraph v BIUA e T?v 田 D2Please answer the following using the experiment data below. Provide the formulas or methods used for calculations of each. Number of colonies on LB/amp/ara plate = Micrograms of DNA spread on the plates = Transformation efficiency = DNA plasmid concentration: 0.08 μg/μl 250 μl CaCl transformation solution 10 μl pGLO plasmid solution 250 μl LB broth 100 μl cells spread on agar 227 colonies of transformants
- 7) Why are the following reagents used? Neutralizing solution (Plasmid isolation) Isopropanol (Plasmid isolation) RNase (isolation of genomic DNA)A target gene for producing human insulin must be inserted into a plasmid before the transformation process. Why the plasmid is important in this process? * I The target gene lacks the ability to replicate itself. II Screening process will be difficult without cloning vector. II The host cell is able to accept the recombinant plasmid but not the non- recombinant plasmid. O I and Il only I and III only O Il and III only O I, Il and IIIDrag and drop the appropriate text in the gaps below. Firstly, the plasmid DNA in the E. coli is propagated through Then the bacterial cells are pelleted by centrifugation at 10,000 RPM. The media supernatant is removed and the bacterial cells are The bacterial cells are then lysed via |. Contaminating macromolecules such as protein and chromosomal DNA is removed by Overnight incubation of the bacterial culture washed in cell resuspension buffer addition of lysis buffer addition of neutralisation buffer washing of the spin column separation via agarose gel electrophoresis