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Team:St Andrews/Procedure
From 2012.igem.org
Lab Book
Procedure
- date
- what we did
- date
- what we did
- date
- what we did
anyone really keen to write this up, up to protein visualization? Don't forget to mention the differing restriction enzymes and vectors of each team. I can add in links to the protocols page.
Please also refer to our Protocols page.
Sequences Primers Sequencing results Lipid analysis
Start from lipid extraction
primers, sequence results
Sequences
atgtcgcgggttgacaatttaactgtcgcgccgggtccaccggatgtcatgaaggcggtgcttaagtctgggcgcaacaaggagcacaat gtcattgtgccttcctccgcactgaccattggagaaattcaggagaaaatcccggtaaagtactttgaacgcaacactacccggagcgtc atgtttcttttacgcgacctcgcacaggtggcgttgacgtacgccattatgtacgccgtcgccttgccacttgccacttccatggaagtttctg cagcaagaacgttggccgacggtggggtgttatcattgatggcaggtacagccatgacgacagcggcgtggcttttaaaggcagtgttgtg ggcggtgttttggtttgtacagggacttaacggcactgcgctttgggtcctggctcacgaatgcggccatcaggcgttcagccccatgaaag gtttgaatgacgccgttggcatgattttgcactctgcgctgctcgtgccgtaccacagctggcgcatcacccacggcggccaccacaaac acacgaatcacctcacgaaggatcttgtgtttgttccagacaaacgaaacgcggttgtggagctcgtggaggaggcgccgctggtgttatt aattcaattattgctgatttttctctttggttggccggcacatcttctttttaatgcctctggacaggaatttggccgactcgcgagccactttgac cccggcgctccatttttccgcagcgaagaccgtcacgatattgtcctgtcgaatgttgggattgtcagcgcgttatttgtcattttctccagcgt ttaccgctttggttttacaaatgttttctgctggtacattgtaccgtacctctgggtgaacttttggcttgtgtacattacatacctgcagcacacg gatatacgcattcctcactacacacatgagcactggacgtttgttcgcggtgcattggcggctgtggacagggactacggctttgtccttaac acatggctccatcacatcaatgattcccacgtggtacatcacctctttagcaaaatgccacactacaacgcaatccaggtgacaagaaag tacattcgtgagatactgggtgccacatacattacggatgagaggtcactgtggaagatgctctgggaacagcgtagagagtgccgctatg ttgttcccgcagagggcgtctgtgtctttcatgggtaa
atgactgccacgattcccccgttgacaccaacggtaacgcccagcaatcccgatcgcccg attgcggatctcaaactacaagacatcattaaaaccctgcccaaggaatgcttcgagaaa aaagcgagcaaagcctgggcttctgttttgattaccctaggggcgatcgccgtgggctat ttgggcattatttatctgccctggtactgcttgcccattacctggatctggacagggaca gccttaacgggggccttcgttgtcggccatgactgtggccatcgctcctttgctaaaaaa cgctgggtcaatgatttagtgggacatatcgcttttgctcccctcatctaccctttccat agctggcgcctactccacgaccaccatcacctccacaccaacaaaattgaggttgataac gcctgggatccctggagtgtggaagctttccaagccagcccggcgatcgtccggcttttt tatcgggccatccggggtcccttctggtggactggttccattttccattggagcttaatg cacttcaaactttccaactttgcccaaagggaccgcaataaagtcaaattatccattgcc gttgtcttcctgtttgcggcgatcgcctttcctgccctaattatcaccacaggggtgtgg ggtttcgtcaaattttggctaatgccctggttggtgtatcacttttggatgagcactttt accattgtgcaccacaccattcccgaaattcgtttccgtcccgccgccgattggagtgcc gccgaagcccagttaaatggtactgttcactgcgattatccccgttgggtggaagtgctc tgccatgacatcaacgtccatattccccaccacctctccgttgccatcccttcctataac ctacgactagcccacggaagtttaaaagaaaactggggaccttttctttacgagcgcacc tttaactggcaattaatgcaacaaattagtgggcaatgtcatttatatgaccccgaacat ggctaccgcaccttcggctccctgaaaaaagtttaa
gtgcgtctagaaatttcatcgcctcaaacaaagcttccttaccccaaaactgaagaatta ccatttaccctccaagagctcagaaacgctattccagcggattgttttgagccatcggta gtccggtccttgggctacttttttttggatgttggtttaattgccgggttttatgctcta gcggcctaccttgattcctggttcttctatccgattttttggttaattcagggaacccta ttctggtccctgtttgtggtgggccatgattgtggccatggctccttttccaaatccaaa acccttaataattggattggtcatctcagccacacgccaattttggtgccttaccatggc tggcgtattagtcatcgtactcaccatgccaacacgggcaatatcgacaccgacgaaagt tggtatccagtgtcggagcaaaaatataaccaaatggcctggtatgaaaaacttctacgt ttttacttgcctctgatcgcctaccccatttatctatttcggcgatcgccaaaccggcaa ggctcccatttcatgcccggcagtcccctattccgtcccggagaaaaagcagctgttctc accagcacctttgcccttgcagcctttgtcggcttccttggctttttaacttggcaattt ggctggctatttttgctgaaattttatgttgccccctacctcgtgtttgtggtgtggtta gatttggtcacatttttacatcacactgaagacaatatcccttggtatcgtggtgatgac tggtattttctcaaaggtgccctctccaccattgatcgggattacggcttcattaacccc attcaccatgacattggcacccacgtcgcccaccatattttctcgaatatgccccactac aagttacgccgggcgactgaagccatcaagcccattttaggggaatattatcgatattct gacgagccaatttggcaagctttttttaagtcctactgggcttgccattttgttcctaat caaggttcaggggtctattaccaatccccatccaatggtggatatcaaaagaaaccttaa
atgctaacagcggaaagaattaaatttacccagaaacgggggtttcgtcgggtactaaac caacgggtggatgcctactttgccgagcatggcctgacccaaagggataatccctccatg tatctgaaaaccctgattattgtgctctggttgttttccgcttgggcctttgtgcttttt gctccagttatttttccggtgcgcctactgggttgtatggttttggcgatcgccttggcg gccttttccttcaatgtcggccacgatgccaaccacaatgcctattcctccaatccccac atcaaccgggttctgggcatgacctacgattttgtcgggttatctagttttctttggcgc tatcgccacaactatttgcaccacacctacaccaatattcttggccatgacgtggaaatc catggagatggcgcagtacgtatgagtcctgaacaagaacatgttggtatttatcgtttc cagcaattttatatttggggtttatatcttttcattcccttttattggtttctctacgat gtctacctagtgcttaataaaggcaaatatcacgaccataaaattcctcctttccagccc ctagaattagctagtttgctagggattaagctattatggctcggctacgttttcggctta cctctggctctgggcttttccattcctgaagtattaattggtgcttcggtaacctatatg acctatggcatcgtggtttgcaccatctttatgctggcccatgtgttggaatcaactgaa tttctcacccccgatggtgaatccggtgccattgatgacgagtgggctatttgccaaatt cgtaccacggccaattttgccaccaataatcccttttggaactggttttgtggcggttta aatcaccaagttacccaccatcttttccccaatatttgtcatattcactatccccaattg gaaaatattattaaggatgtttgccaagagtttggtgtggaatataaagtttatcccacc ttcaaagcggcgatcgcctctaactatcgctggctagaggccatgggcaaagcatcgtga
atgaaggtcatcgtcgcttctggcccggacggtgcccgcaagcacgaggtggagctggca gccaacgccacgctcgcagatctgaagaaggcctaccaacggggtgtggacgtgcaccgc aagtcgttcaaggttcccagcgcggagtcgccgctgccaggtgcggatagtggcaagctg cgcccgaacctcattactctgtcagataaggtgcccctgtcgcagcagggggtgaaggat ggctcggtgatcacttacaaggacctcggcccgcagatcggctaccgcacggtgttctac gtcgagtatgccggccccatcgccttcatgctgctgtacgccatgcgcccttcgctcatc tacggctctgccccgatgccggcttacggctacacgcagaagctatacattggcctcttc ctcgcccacttcttaaagcgcgagctcgagtccatgtttgtgcacaagttctcgcaccca acgatgccgatgcgcaacatcttcaagaactgcatctactactggtccttcgccgccttc atcggctacgtgctgtgcagtccttcattcacgccgaccagcaccatgcagtcaaacttc ggcgccgtggtcatggtcatcaacgagctgctgaacttcgcggtgcactaccagcttagc acgatgcgcaagtccgatggtgacaccacccgcaacgtgccgcaaggccctctgttcgcc ttcgtctcgtgcccgaactacttctttgagattatgtcgtgggtgtccttttccatcggc acaaatatgttatcctcctggttcttcacactcgccggtttcgtgcagatggcggactgg gcgaagaagaagcaccggggctacgtcaaggcggacccggccaataaaaagaaggccgcc attctgcccttcatcatgtag
Primers
All primers are notated 5' to 3'.
When using the pET-duet vector, we needed additionally primers for the alternative restriction sites and His-tags.
Sequencing results
- date and gene
- result
- date and gene
- result
- date and gene
- result
- 23/7/12 Δ12 Syn.
15b forward gave 0 nucleotides
15b reverse
20b forward gave a 41 nucleotide result.
SequencegCGnCctGntGCCgCGCGGCnnnnnnnnnnnnnnnnnngacA BLAST search showed this to be compatible with Crocosphaera watsonii (genome shotgun sequence, 21%), a diazotrophic cyanobacteria. Add in relations to synechocystis.
20b reverse
Lipid analysis
who knows what those Metal Mickies are up to. Eating nachos in the Whey Pat most likely.
