a. The recognizing sequence is the restriction site where the restriction enzyme recognize the site and cleave the restriction fragments. the sequence may vary based upon the enzyme.
Ndel sequence is: 5'CATATG3'
3'GTATAC5'
BamH1 sequence is: 5'GGATCC
3'CCTAGG
b.) PCR is a polymerize chain reaction where it needs a pair of primers to amplify the given template and resulting in the formation of product.
Forward primer: GCCTCCGGTAAGTCCGGCCGCT
Reverse primer :CGCGGCAGCGCGCGAGTGGCCGA
melting temperature can be estimated by formula: Tm = 4(G+C)/2(A+T)
1) Imagine that you are interested in studying the enzyme dihydrofolate reductase (DHFR) from Mycobacterium tuberculosis...
3. (2 pts.) You are sequencing the end of a genomic DNA fragment that was cut with the restriction enzyme BamH1 and ligated into a plasmid that was also cut with BamH1. You have denatured the DNA and annealed a labeled primer to plasmid sequences adjacent t<o the insertion site as shown below: 3' BamHi 5' GATCTAGCTAGCTAGCTAGCTAGCTAGCTAGCCATCGATGCTAGGAATCTTTGCTGATGCTAGTCGATGCCGTAGC ACTACGATCAGCTACGGC 3' 18 base primer 5' Next you add DNA polymerase, buffer, an excess of all four dNTPs, and a small amount of...
Suppose you digest the genomic DNA of a particular organism with the restriction enzyme SauA. Then you ligate the resulting fragment into a unique BamHI cloning site of a plasmid. The sequence of the restriction sites and position of cleavage is shown below Note: X and Y and their complementary bases Z and Y’ respectively can be any base (A,C, G, or T) 1) As you can see, ligation is possible because the two restriction enzymes produce compatible sticky ends....
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...
Below is a set of experiments for cloning the human growth hormone (HGH) gene and then expressing the HGH protein in E. coli starting from human pituitary glands and a tube of plasmid vector DNA. The plasmid expression vector contains the arabinose inducible expression system. Specify the correct order for carrying out these experiments, using the letter codes in front of each procedure. Use all of the steps in your answer, except for one step that should NOT be included....
These are previous exam questions I am just preparing for the final exam by studying the previous questions! please help me out! A1. The following is the DNA sequence of a hypothetical gene for the SMALL protein. It is called the SMALL gene. 1 atgggattac actgtcacga ccaaatagcc ttcattgtat 41 caaaaggatc aatcgagtta tag Imagine you are doing a research project in a laboratory and your supervisor asks you to clone the SMALL gene into the pBR322 plasmid (shown below) You must...
I didnt do quite as well as i wpuld have liked on my molecular bio exam. Please help me reciew what I missed. Thanks! We were unable to transcribe this imageWe were unable to transcribe this imageA fragment restricted with EcoRI enzyme can be used for ligation into a plasmid that was restricted with BamHI because both the insert and the plasmid contain sticky ends a True b) False 10. Polynucleotide probes can be used to screen a genomic library...
In your previous prac session you digested your POTC-A plasmid DNA with 3 enzyme mixes (AB and C). One mix contained the restriction endonuclease Kpnl alone, another contained both del and Not and another contained Sphi and Nhe, but you don't know which was which yet. The sites where these enzymes cut POTC-A are indicated on the plasmid map at the top of the page. Calculate the sizes of the DNA fragments that you would expect to see on your...
1) You have two human liver cells (A and B) and you hypothesize that the insulin receptor gene in Cell A has a mutation in exon 1 and Cell B contains the wild type sequence. You extract genomic DNA from each of the cells. Of the following, what would be the most efficient (quick, precise and relatively cheap) way to test your hypothesis. a. Isolate protein from both cells, purify the insulin receptor, and determine the amino acid content. b. Sequence the...
Draw (or arrange) only the items you need from the Lab Materials Page (picture below) in the order you would use them to make AND screen a gene library. You will ‘draw’ the steps that use the Lab Materials in the correct order to create and screen a gene library which has the assigned gene clone. You will need to add some connecting words and phrases to make all the steps for making your library clear. The genomic gene clone...
A1. The following is the DNA sequence of a hypothetical gene for the SMALL protein. It is called the SMALL gene. i atgggattac actgtcacga ccaaatagcc ttcattgtat 41 caaaaggato aatcgagtta tag Imagine you are doing a research project in a laboratory and your supervisor asks you to clone the SMALL gene into the PBR322 plasmid (shown below). You must use the Pstl and EcoRI sites for your cloning. HindIII EcoRI | EcoRV BamHI 4359 0 29 185 4000 375 Sall Psti...