Jessica's final year project deals with the metagenomic studies of anfH homologous genes from freshwater lake samples in Kampar Eastlake. She successfully cloned and sequenced two partial anfH homologous gene sequences. BlastX analyses revealed that one of the DNA sequences showed highest similarity to AnfH protein from a Brazilian soil bacteria isolate while the second sequence showed highest identity to the AnfH protein sequence of a bacterium that was isolated from Tonle Sap, a freshwater lake in Cambodia. To determine their evolutionary relationship, she subsequently constructed a phylogenetic tree using the corresponding protein sequences. Describe the steps that are required to perform the analysis.
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- TRANSCRIBE this DNA sequence: TACGTTACT AUGCAAUGA O ATGCAATGA AUGGATUGA TACGTTACTexplain how the following genotypic andphenotypic methods of microbe identification is done. Use diagrams or schematic representations to explain yourwork. (1) Restriction Fragment Length Polymorphism (RFLP)(2) Pulsed-Field Gel Electrophoresis (PFGE)(3) Whole Genome Sequencing (WGS)(4) Gram Staining(5) Biochemical Reactions: Multiple Tube Fermentation Technique(6) Biochemical Reactions: IMViC TestWhat key molecules are essential for Sanger sequencing?
- Nicotinamide Adenine Dinucleotide (NAD) Assay (SIGMA Kit MAK037) analysis of tissue samples initially requires: centrifugation at 13,000g for 10min to homogenise tissue. centrifugation of 20mg of tissue at 2,000rpm with Extraction Buffer. freezing and thawing for 2 cycles of 20min before addition of Extraction Buffer. homogenization of PBS washed tissue with Extraction Buffer.RESEARCH ARTICLE TITLE: "On the rapidity of antibiotic resistance evolution facilitated by a concentration gradient" Full Article Reseach: Hermsen, R., Deris, J. B., & Hwa, T. (2012). On the rapidity of antibiotic resistance evolution facilitated by a concentration gradient. Proceedings of the National Academy of Sciences, 109(27), 10775–10780. doi:10.1073/pnas.1117716109 QUESTION: 1. In biological/science manner, what are your comments on how the authors constructed their ARTICLE/RESEACH TITLE (see article).Q2 (a) In 2014, during their iGEM project on biodesalination, students were willing to modify bacteria so that instead of swimming they would float. To do this, they chose to first knock-out one of the genes for motility (such as MotA) and then insert a gene sequence to make gas bubbles. They used the restriction enzyme Pstl, which recognizes the sequence 5'-CTGCAG-3' and cuts between A-G. (CTGCA/G). Using the sequence from one of the DNA strands of the gene below, 5'-TTGATTTTATCGCTGCAGTCGATTATCTGCGCCTGATTATCACTGCAGGCG-3' (fragment modified from the students' plasmid) (v) Provide the double strand sequences resulting from the action of Pstl. (b) (vi) (vii) Specify the name of the ends of the sequences generated and explain their functionality. Describe the last step required to complete the process of deletion started using Pstl (viii) Provide the number of amino-acids deleted through this process and explain why such a small deletion could be sufficient to inactivate motility. The…
- Q2 (a) In 2014, during their iGEM project on biodesalination, students were willing to modify bacteria so that instead of swimming they would float. To do this, they chose to first knock-out one of the genes for motility (such as MotA) and then insert a gene sequence te make gas bubbles. They used the restriction enzyme Pstl, which recognizes the sequence 5'-CTGCAG-3' and cuts between A-G. (CTGCA/G). Using the sequence from one of the DNA strands of the gene below, 5'-TTGATTTTATCGCTGCAGTCGATTATCTGCGCCTGATTATCACTGCAGGCG-3' (fragment modified from the students' plasmid) (v) Provide the double strand sequences resulting from the action of Pstl.2b)An ancient gene underwent duplication during the course of evolution to yield two chain genes designated as δ and ε genes. Jessica’s final year project deals with the metagenomic studies of anfH homologous genes from freshwater lake samples in Kampar Eastlake. She successfully cloned and sequenced two partial anfH homologous gene sequences.BlastX analyses revealed that one of the DNA sequences showed highestsimilarity to AnfH protein from a Brazilian soil bacteria isolate while the second sequence showed highest identity to the AnfH protein sequence of a bacterium that was isolated from Tonle Sap, a freshwater lake in Cambodia. To determine their evolutionary relationship, she subsequently constructed a phylogenetic tree using the corresponding protein sequences. Describe the steps that are required toperform the analysis.When Griffith injected mice with a combination of live rough-strain and heat-killed smooth-strain pneumococci, he discovered that (a) the mice were unharmed (b) the dead mice contained living rough-strain bacteria (c) the dead mice contained living smooth-strain bacteria (d) DNA had beentransferred from the smooth-strain bacteria to the mice (e) DNA had been transferred from the rough-strain bacteria to the smooth-strain bacteria
- How to isolate cpDNA for whole chloroplast DNA sequencing (MiSeq system)? Any universal primer or cpDNA extraction kit support for this?Traditional Sanger sequencing has largely been replaced in recent years by next-generation and third-generation sequencing approaches. Describe advantages of these sequencing methods over first-generation Sanger sequencing.does Sanger sequencing still used?