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Interesting Tuur
- 1. YOGHURT
- 2. Lactobacillus Delbrueckii subsp. Bulgaricus Streptococcus Salivarius subsp. Thermophilus
- 4. LACTOBACILLUS TRANSFORMATION Estimated time for procedure: 3-4 days CELL CULTURE ELECTRICAL PULSE Inoculate serial dilutions of fresh bacterial culture into 100 ml of Mix 80 µl of cell suspension with 0.3 to 2 µg of plasmid DNA MRS containing 0.1% glycine and incubate at 42°C overnight. Subject sample to a 1-kV, 800-Ω, 25-µF electric pulse in a 0.2-cm Harvest 10 ml of culture cells at beginning of stationary phase cuvette by using a Gene Pulser and a Pulse Controller apparatus. (optical density at 600 nm, 1.7) by centrifugation (need ~2.3mL of culture per electroporation) EXPRESSION Immediately add 2 milliliters of milk medium (0.2 M sucrose, 5% WASH BUFFER skim milk, 0.1% yeast extract, 1% Casamino Acids, 25mM `Wash bacteria once with 100 ml of cold electroporation buffer MgCl2) (EB) (0.4 M sucrose, 1 mM MgCl2, 5 mM Kh2PO4; PH 6) Wash bacteria twice with 30 ml of cold EB PLATING, SELECTION Incubate cells for 3h at 37°C before plating on MRS agar THERMAL SHOCK supplemented with antibiotics. Add antibiotic erythromycin at concentration 7.5 µg/ml and chloramphenicol at concentration Resuspend cells in EB to an optical density at 600 nm of about 50 7.5 µg/ml depending on plasmid resistance gene Incubate cell suspension at 45°C for 20 min then keep on ice for Incubate plates at 37°C for 2 to 3 days under anaerobic 10 min conditions in jars containing GasPak
- 5. CELL CULTURE
- 6. INCUBATE
- 14. CENTRIFUGE
- 19. local water
- 20. London biotopes
- 23. TCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACAT TAAAGAGGAGAAAATGCCTCTTCCGCTGTACCTGCTCGCCGTGGCCGTCTGCG CCATGGGCACCTCGGAGTTCATGCTCGCCGGTCTCGTGCCGGACATCGCCTCG GATCTCGGCGTCACCGTCGGGACCGCAGGCACGCTCACCTCGGCCTTCGCGA TCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACAT CCGGCATGATCGTCGGCGCTCCCCTCGTGGCGGCGCTGGCCCGCACCTGGCC TAAAGAGGAGAAAATGAGTATTCAACATTTTCGTGTCGCCCTTATTCCCTTTTTTGC CAGGCGTTCCAGCCTCCTCGGATTCATCCTCGCCTTCGCGGCGGCACACGCCG GGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGAT TGGGAGCCGGCACCACGAGCTTCCCCGTCCTGGTGGCCTGCCGGGTCGTGG GCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAG CCGCGCTCGCGAACGCGGGATTCCTCGCGGTCGCACTGACGACTGCCGCCG CGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACT CACTGGTCCCTGCCGACAAGAAGGGACGCGCGCTGGCCGTGCTGCTGTCCG TTTAAAGTTCTGCTATGTGGTGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGC GCACGACGGTGGCCACGGTCGCCGGCGTCCCCGGCGGGTCACTCCTCGGCA AACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGT CATGGCTCGGCTGGCGGGCCACGTTCTGGGCCGTCGCCGTCTGCTGCCTGCC CACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTG CGCGGCGTTCGGCGTGCTGAAGGCAATCCCCGCCGGACGTGCGACGGCAG CCATAACCATGAGTGATAACACTGCTGCCAACTTACTTCTGACAACGATCGGAG CGGCGACCGGTGGGCCGCCGCTGCGAGTCGAGCTCGCCGCGCTCAAGACC GACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGC CCCCGGTTGCTGCTGGCGATGCTGCTGGGCGCGCTGGTGAACGCGGCAACCT CTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTG TCGCGAGCTTCACCTTCCTGGCCCCCGTCGTGACCGACACCGCAGGGCTGGG ACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGC CGACCTGTGGATCTCTGTCGCCCTGGTGCTCTTCGGCGCCGGTTCCTTCGCCGG GAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGAT CGTCACCGTCGCCGGACGACTGTCCGACCGACGCCCCGCCCAGGTGCTCGCC AAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCT GTCGCCGGTCCGCTGCTGCTCGTCGGCTGGCCCGCGCTGGCGATGCTGGCCG GATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGG ACCGGCCGGTCGCCCTGCTGACCCTCGTGTTCGTCCAAGGCGCACTGTCGTTC GCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG GCGCTGGGCAGCACGCTGATCACGCGGGTCCTCTACGAGGCGGCGGGAGC CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTA ACCCACCATGGCCGGTTCGTACGCGACCGCCGCCCTCAACGTGGGCGCCGC AGCATTGGTAAATTAAAGAGGAGAAAATGGCTTCCTCCGAAGACGTTATCAAA GGCCGGACCGCTCGTCGCCGCGACCACTCTCGGCCACACGACCGGCAACCT GAGTTCATGCGTTTCAAAGTTCGTATGGAAGGTTCCGTTAACGGTCACGAGTTCG CGGGCCGCTGTGGGCGAGCGGGCTCCTGGTCGCCGTCGCGCTGCTCGTCGC AAATCGAAGGTGAAGGTGAAGGTCGTCCGTACGAAGGTACCCAGACCGCTAA GTTCCCCTTCCGCACGGTGATCACGACGGCCGCACCCGCCGACGCGACCCG ACTGAAAGTTACCAAAGGTGGTCCGCTGCCGTTCGCTTGGGACATCCTGTCCCC GTGAATTAAAGAGGAGAAAATGCGTAAAGGAGAAGAACTTTTCACTGGAGTTG GCAGTTCCAGTACGGTTCCAAAGCTTACGTTAAACACCCGGCTGACATCCCGG TCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTG ACTACCTGAAACTGTCCTTCCCGGAAGGTTTCAAATGGGAACGTGTTATGAACTT GAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCA CGAAGACGGTGGTGTTGTTACCGTTACCCAGGACTCCTCCCTGCAAGACGGTG CTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCGGTTATGGT AGTTCATCTACAAAGTTAAACTGCGTGGTACCAACTTCCCGTCCGACGGTCCGG GTTCAATGCTTTGCGAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGA TTATGCAGAAAAAAACCATGGGTTGGGAAGCTTCCACCGAACGTATGTACCCG GTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACG GAAGACGGTGCTCTGAAAGGTGAAATCAAAATGCGTCTGAAACTGAAAGACG GGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATA GTGGTCACTACGACGCTGAAGTTAAAACCACCTACATGGCTAAAAAACCGGTT GAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAA CAGCTGCCGGGTGCTTACAAAACCGACATCAAACTGGACATCACCTCCCACAA ATTGGAATACAACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAA CGAAGACTACACCATCGTTGAACAGTACGAACGTGCTGAAGGTCGTCACTCCA GAATGGAATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCGT CCGGTGCTTAATAATATAAACGCAGAAAGGCCCACCCGAAGGTGAGCCAGTG TCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTT TGACTCTAGTAGAGAGCGTTCACCGACAAACAACAGATAAAACGAAAGGCCC TTACCAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGATCCCAACG AGTCTTTCGACTGAGCCTTTCGTTTTATTTGATGCCTGG AAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGCTGCTGGGATTACAC ATGGCATGGATGAACTATACAAATAATAATATAAACGCAGAAAGGCCCACCCG AAGGTGAGCCAGTGTGACTCTAGTAGAGAGCGTTCACCGACAAACAACAGATA AAACGAAAGGCCCAGTCTTTCGACTGAGCCTTTCGTTTTATTTGATGCCTGG
- 24. HARVEST
- 25. thank you