Recitation 8 - Spring 2022 B The Holechek lab has sequenced a bacterial gene (LacZ) coding for a protein (ß-galactosidase) that breaks down lactose into glucose and galactose. The sequence for the non-coding strand is as follows below, with spaces between every 10 base pairs: 5'-TTTTTGA CACCAGACCA ACTGGTAATG GTAGCGACCG GCGCTCAGCT GGAATTCCGC CGATACTGAC GGGCTCCAGG AGTCGTCGCC ACCAATCCCC ATATGGAAAC CGTCGATATT CAGCCATGTG CCTTCTTCCG CGTGCAGCAG ATGGCGATGG CTGGTTTCCA TCAGTTGCTG TTGACTGTAG CGGCTGATGT TGAACTGGAA GTCGCCGCGC CACTGGTGTG GGCCATAATT CAATTCGCGC GTCCCGCAGC GCAGACCGTT TTCGCTCGGG AAGACGTACG GGGTATACAT GTCTGACAAT GGCAGATCCC AGCGGTCAAA ACAGGCGGCA GTAAGGCGGT CGGGATAGTT TTCTTGCGGC CCTAATCCGA GCCAGTTTAC CCGCTCTGCT ACCTGCGCCA GCTGGCAGTT CAGGCCAATC CGCGCCGGAT GCGGTGTATC GCTCGCCACT TCAACATCAA CGGTAATCGC CATTTGACCA CTACCATCAA TCCGGTAGGT TTTCCGGCTG ATAAATAAGG TTTTCCCCTG ATGCTGCCAC GCGTGAGCGG TCGTAATCAG CACCGCATCA GCAAGTGTAT CTGCCGTGCA CTGCAACAAC GCTGCTTCGG CCTGGTAATG GCCCGCCGCC TTCCAGCGTT CGACCCAGGC GTTAGGGTCA ATGCGGGTCG CTTCACTTAC GCCAATGTCG TTATCCAGCG GTGCACGGGT GAACTGATCG CGCAGCGGCG TCAGCAGTTG TTTTTTATCG CCAATCCACA TCTGTGAAAG AAAGCCTGAC TGGCGGTTAA ATTGCCAACG CTTATTACCC AGCTCGATGC AAAAATCCAT TTCGCTGGTG GTCAGATGCG GGATGGCGTG GGACGCGGCG GGGAGCGTCA CACTGAGGTT TTCCGCCAGA CGCCACTGCT GCCAGGCGCT GATGTGCCCG. GCTTCTGACC ATGCGGTCGC GTTCGGTTGC ACTACGCGTA CTGTGAGCCA GAGTTGCCCG GCGCTCTCCG GCTGCGGTAG TTCAGGCAGT TCAATCAACT GTTTACCTTG TGGAGCGACA TCCAGAGGCA CTTCACCGCT -3' 1) What "mystery species" does our sequence come from? Use the NCBI Nucleotide blast from the website https://blast.ncbi.nlm.nih.gov/Blast.cgi to identify the bacterial species this gene belongs to. Copy ONLY the sequence into the blast, do not include the 5'- or -3', and leave the spaces between every 10 nucleotides. Organism's scientific name: Escherichia coli 2) Here is one set of primers used to amplify a segment of this gene. Where does each primer anneal (bind)? Highlight, underline or put a box around the annealing sites for the FORWARD and REVERSE primers on the sequence above (REMEMBER! Reverse primer sequences bind to the coding strand.) *HINT* One primer will be found within each bolded region. Forward primer sequence: 5'- TCTGGATGTCGCTCCACAAA -3' Reverse primer sequence: 5'- CCAACTGGTAATGGTAGCGA -3' вать ассаттассат сост 3) Find the proper annealing temperature for the forward and reverse primer from the Short Answer using this website: https://tmcalculator.neb.com/#!/main (Links to an external site.). (leave first 3 selections alone, assume product group Q5, polymerase Q5 high-fidelity DNA polymerase, and 500nM primer concentration) ( 66 62 Forward Primer Temperature: Reverse Primer Temperature:

Recitation 8 - Spring 2022 B The Holechek lab has sequenced a bacterial gene (LacZ) coding for a protein (ß-galactosidase) that breaks down lactose into glucose and galactose. The sequence for the non-coding strand is as follows below, with spaces between every 10 base pairs: 5'-TTTTTGA CACCAGACCA ACTGGTAATG GTAGCGACCG GCGCTCAGCT GGAATTCCGC CGATACTGAC GGGCTCCAGG AGTCGTCGCC ACCAATCCCC ATATGGAAAC CGTCGATATT CAGCCATGTG CCTTCTTCCG CGTGCAGCAG ATGGCGATGG CTGGTTTCCA TCAGTTGCTG TTGACTGTAG CGGCTGATGT TGAACTGGAA GTCGCCGCGC CACTGGTGTG GGCCATAATT CAATTCGCGC GTCCCGCAGC GCAGACCGTT TTCGCTCGGG AAGACGTACG GGGTATACAT GTCTGACAAT GGCAGATCCC AGCGGTCAAA ACAGGCGGCA GTAAGGCGGT CGGGATAGTT TTCTTGCGGC CCTAATCCGA GCCAGTTTAC CCGCTCTGCT ACCTGCGCCA GCTGGCAGTT CAGGCCAATC CGCGCCGGAT GCGGTGTATC GCTCGCCACT TCAACATCAA CGGTAATCGC CATTTGACCA CTACCATCAA TCCGGTAGGT TTTCCGGCTG ATAAATAAGG TTTTCCCCTG ATGCTGCCAC GCGTGAGCGG TCGTAATCAG CACCGCATCA GCAAGTGTAT CTGCCGTGCA CTGCAACAAC GCTGCTTCGG CCTGGTAATG GCCCGCCGCC TTCCAGCGTT CGACCCAGGC GTTAGGGTCA ATGCGGGTCG CTTCACTTAC GCCAATGTCG TTATCCAGCG GTGCACGGGT GAACTGATCG CGCAGCGGCG TCAGCAGTTG TTTTTTATCG CCAATCCACA TCTGTGAAAG AAAGCCTGAC TGGCGGTTAA ATTGCCAACG CTTATTACCC AGCTCGATGC AAAAATCCAT TTCGCTGGTG GTCAGATGCG GGATGGCGTG GGACGCGGCG GGGAGCGTCA CACTGAGGTT TTCCGCCAGA CGCCACTGCT GCCAGGCGCT GATGTGCCCG. GCTTCTGACC ATGCGGTCGC GTTCGGTTGC ACTACGCGTA CTGTGAGCCA GAGTTGCCCG GCGCTCTCCG GCTGCGGTAG TTCAGGCAGT TCAATCAACT GTTTACCTTG TGGAGCGACA TCCAGAGGCA CTTCACCGCT -3' 1) What "mystery species" does our sequence come from? Use the NCBI Nucleotide blast from the website https://blast.ncbi.nlm.nih.gov/Blast.cgi to identify the bacterial species this gene belongs to. Copy ONLY the sequence into the blast, do not include the 5'- or -3', and leave the spaces between every 10 nucleotides. Organism's scientific name: Escherichia coli 2) Here is one set of primers used to amplify a segment of this gene. Where does each primer anneal (bind)? Highlight, underline or put a box around the annealing sites for the FORWARD and REVERSE primers on the sequence above (REMEMBER! Reverse primer sequences bind to the coding strand.) HINT One primer will be found within each bolded region. Forward primer sequence: 5'- TCTGGATGTCGCTCCACAAA -3' Reverse primer sequence: 5'- CCAACTGGTAATGGTAGCGA -3' вать ассаттассат сост 3) Find the proper annealing temperature for the forward and reverse primer from the Short Answer using this website: https://tmcalculator.neb.com/#!/main (Links to an external site.). (leave first 3 selections alone, assume product group Q5, polymerase Q5 high-fidelity DNA polymerase, and 500nM primer concentration) ( 66 62 Forward Primer Temperature: Reverse Primer Temperature:

Biology 2e

2nd Edition

ISBN:9781947172517

Author:Matthew Douglas, Jung Choi, Mary Ann Clark

Publisher:Matthew Douglas, Jung Choi, Mary Ann Clark

Chapter14: Dna Structure And Function

Section: Chapter Questions

Problem 26CTQ: Prokaryotes have a single circular chromosome while eukaryotes have linear chromosomes. Describe one...

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