Sprint 9:53 PM BIOL101 Spring2016 Final.pf Done Molecular Biology 10f 4 ~2016 In
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Sprint 9:53 PM BIOL101 Spring2016 Final.pf Done Molecular Biology 10f 4 ~2016 Instructions Write your name on ALl pages Restrict your answers to the space and word limits provided or irrelevant indormation will resu in a loss of points 1J Primer Extension and labeling (9 points You are setting up 3 primer extension reactions wih all the correct butfers and temperatures. For every reaction, draw your primer annealing to the ssONA templabe, the direction of primer extension, and the banding pattem you expect the reactionto produce. ·ndcates radiolabel that does notinterfere with reactin Every Reaction DNA mplate: DNA polymerasedATETP dOT Reaction 1 Draw annealing of priner and direction of primer extenaion Deaw annealing of priner and dizection of pelmer extenslon Deaw aealing of primer and direction of prime excenalon 2.1 Retroviruses Retroviruses can insert DNA randomly into the human genome. Depending on its location this integration can have condequences ol varying severty. Describe 3 general integration scenaries. (where does the retroviral DNA integrabe? What are the comequences?) (word limit: 6o) [9 points) 3.) RNA secondary structure Due to its single stranded nature RNA can basepair with itself and form secondary structures like stem loops Desoribe 3 examples of RNA secondary structures and their function [Word limit 601 (9 points) Biol 201, Spring 2036 Final Exam, June 4.) Southern ?1ots You run a Southern Blot using penomic DNA and a probe aligning to the piece of DNA 00 00 800 9000 1200 1400 1800 1800 2000 ?Explanation / Answer
Reaction 1 primer orientation is wrongly given, here there wont be any extension because polymerase extends from 5’- to 3’ direction on primer 3’ side template is absend hence cannot extend
3’-TATAGCGATGCAGGTCAAATGC-5’
Template 5’-ATATCGCTACGTCCAGTTTACGtcaagttgcacacgtgcaga-3’
22 base pairs appear in the gel at 22nd position in lane 1 (you may draw a line/bar on the side diagram)
Reaction 2
Primer 3’-GCAGTTCAACGTGTGCACGTCT-5’
Template 5’-atatcgctacgtccagtttaCGTCAAGTTGCACACGTGCAGA-3’
42 base pairs appear in the gel at 42nd position in lane 2
Reaction 3
Primer 3’-GCAGTTCAACGTGTGCACGTCT-5’
Template 5’-atatcgctacgtccagtttaCGTCAAGTTGCACACGTGCAGA-3’
42 base pairs appear in the gel at 42nd position in lane 3
2. Retroviruses not completely random, however they randomly integrate in to transcription active sites. Onco retroviruses like MLV and ASLV integrate in transcriptional start sites and deregulates gene expression. Results in insertional activation of proto oncogenes. For example with an ASLV MLV-based vector delivering the IL2RG gene developed T cell leukemia. HIV prefers integration in transcription units may result in cancer.
3. Rna Double helix resembles A form of DNA and patterns of double helix RNA is volved in recognition sites for vareous translation factors and proteins. Stem loop structure in tRNAs help in recognising ribosomal binding sites. Pseudoknots RNA of the telomerase enzyme involved in catalytic function
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