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- Using the “target DNA” sequence provided in the attached image, what sizes of inserts can be prepared using HindIII digestion? Which one contains the coding sequence?Restriction digestion and Gel electrophoresis: A single strand of a double-stranded DNA sequence is shown below. Draw a complementary DNA strand and show the restriction digestion pattern of the double-stranded DNA with BamHI and Pst1. Show the separation pattern of the undigested and the digested DNA on your agarose gel. Label the gel appropriately. 5’ – CGAGCATTTGGATCCTGTGCAATCTGCAGTGCGAT – 3’RESTRICTION ENZYME MAPPING Predict the result
- b. Cleavage with chymotrypsin produces the following fragments: Band A: CN , NLQY, GIVEQCCHKRSEY Band B: F, Y, DPTKM, IACCVRGF, RTTGHLCGKDLVNALY Cleavage with Staphilococcus aureus V8 protease produces the following fragments: Band A: GIVE, YNLQNYCN, QCCHKRCSE Band B: PTKM, RTTGHLCGKD, LVNALYIACGVRGFFYD What is the amino acid sequence of the protein? Type your responseUse the sequence provided and make use of figure 1 to determine what restriction enzyme uses the spesific recognition site and figure 2 and 3 to detremine how many times does the sequence occur in the λ DNA sequence? GGATCC: __________________________________________________________ GAATTC: ___________________________________________________________ AAGCTT: ___________________________________________________________Anglais Name and describe in detail three (3) disadvantages of differential centrifugation
- a)List and describe the purpose of each component of the restriction enzyme digest. b)Why do some tubes have no restriction enzyme?Name 4 steps that you can use to isolate a protein from a liver homogenate. Also, briefly discuss each method/step so that the basic principle of each step is clear.Why is it not advisable to use adhesive mixtures if protein histological inevstigations are contemplated.
- Write down a detailed description about the formation of polyacrylamide gel, how it is used for protein separations and the process of electrophoresis in these gels.Discuss/explain/describe in detail the labeling reaction used to label your probe.The same restriction digestion experiment was repeated for EcoR1 on the same DNA molecule. However, the incubation period of the reaction was lengthened for an additional 2 hours due to some unforeseen circumstances. Illustrate below the results you would observe after gel electrophoresis and explain your findings.