Which of the following sequences, in the double-stranded form, can probably be recognized by a restriction enzyme?   Choose an answer from below? TTCCTT CGACGA GTCGAC GGTTGG GAGGAC

Restriction endonucleases are bacterial enzymes that cleave duplex (double-stranded) DNA at specific nucleotide sequences. The mode of replication of the animal virus SV40 has been investigated by using restriction endonucleases that cleave SV40 DNA into a number of unique segments. Like most viruses, SV40 DNA is circular. The map positions of the 11 fragments produced by a pair of restriction endonucleases are shown on the next page. Immediately following a 5 or 10 minute pulse of radioactively labeled thymidine, labeled SV40 molecules that have completed replication during the pulse are isolated. These newly replicated DNA molecules are digested by the restriction endonucleases and the resulting fragments are analyzed for the relative amounts of pulse label they contain. The results are in the table below. Assume that at the time the label was added there was a random population of replicating SV40 DNA molecules in all possible stages of synthesis. From the information given below,…

A linear DNA fragment was produced by digestion with the restriction enzyme, Xba1. This fragment with XbaI(X) sites on both ends was then further digested with HindIII (H) and EcoRI (E).  Draw a restriction map of the linear fragment based on the gel electrophoresis results shown below.  X         H         Marker             E          H/E                           __2000bp __                    __1500b                         __1300bp        __             __        __1000bp                    __                         __700bp             ­­__        __500bp                         __400bp                         __300bp                      __                         __200bp          __        __                         __100bp

The most usefulplasmids contain a _______, which is a short, syntheticDNA sequence that contains a number of different restriction enzyme restriction sites

How many possible restriction maps are compatible with these data?

Consider the ends of the DNA fragments shown below. They have been produced by digestion of a single sequence of DNA using a number of restriction endonucleases. 1. 5'A         3' 3'TTCGA5' 2. 5'G          3' 3'CAGCT5'     3. 5'AATTC3' 3'         G5 4. 5'TCGAC3' 3'         G5'     5. 5'GGG 3' 3'CCC 5'   Which of these ends are capable of annealing and being joined by DNA ligase?

Now that you’ve isolated the gene and made lots of copies, you need to insert the gene into something you can manipulate and move into your lab strain of E. coli.  You have access to the following vectors, either pBR322 or pUC19 (look them up here to see a map https://www.neb.com/products/dna-plasmids-and-substrates Please note the restriction sites in BOLD appear only once in the plasmid).  You may use either vector.  What is a vector? Which vector did you choose? Explain why you made that choice. What enzyme(s) will you use to place your fragment into the vector? Explain why you chose these enzymes. How will you move this construct into coli? What phenotype will tell you if the coli have taken up the plasmid? What phenotype will tell you if the coli have taken up the plasmid with the gasP gene?

Use the plasmid map for pBashi below to answer the following questions Xhol Hind!!! 15 kb 11 kb 10 kb 8 kb 7 kb 5 kb a. How large is pBashi? b. On the gel below, draw the bands corresponding to the expected pBashi cleavage products after restriction digest with the indicated restriction enzymes (Lane 2 - Xhol only, Lane 3 - HindIII only, Lane 4 - Xhol+ HindIII) 4 kb 3 kb 3 kb 2 kb 1 kb 4 kb Hindill 1 kb 7 kb Xhol Xhol Xhol Xhol Hind!!! Ladder Xhol Hindill Hind!!! Pstl Pstl c. Lane 5 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ Pstl. Lane 6 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ HindIII + Pstl. Indicate your final answers to the questions below clearly on the pBashi plasmid above; you will NOT receive any credit if you only list sizes of fragments below. If you need to, draft your answer somewhere else before writing your final answer above.…