Team:Yale/Our Project/Notebook/Week 2

From 2010.igem.org

(Difference between revisions)
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<li>It’s unclear whether the cells at the higher copper concentrations die or simply stop growing.  Spot a drop from each well of the plate onto a LB agar plate and incubate overnight at 37˚C.  Will check for growth to determine survival of different strains at different copper concentrations.</li>
<li>It’s unclear whether the cells at the higher copper concentrations die or simply stop growing.  Spot a drop from each well of the plate onto a LB agar plate and incubate overnight at 37˚C.  Will check for growth to determine survival of different strains at different copper concentrations.</li>
</ul>
</ul>
 +
<i>This information is also recorded on page 11 of the hard copy lab notebook. </i> <br/>
<h4> Plasmid Ligation Strategy</h4>
<h4> Plasmid Ligation Strategy</h4>
<b> The Plan for phs plasmid creation </b> <br/>
<b> The Plan for phs plasmid creation </b> <br/>
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C-terminal primer for phsABC: 5’ AGC ACTAGT A TCA GAC GGC GGA CTT ATC CCC <br/>
C-terminal primer for phsABC: 5’ AGC ACTAGT A TCA GAC GGC GGA CTT ATC CCC <br/>
-
<a id="link" href="javascript:ReverseDisplay('sequence')">See/hide sequence data</a>
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<a id="link" href="javascript:ReverseDisplay('sequence')">See/hide phsABC sequence </a>
<div style="display:none;" id="sequence">
<div style="display:none;" id="sequence">
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phsA STM2065 <br/>
phsA STM2065 <br/>
>sp|P37600|PHSA_SALTY Thiosulfate reductase OS=Salmonella typhimurium GN=phsA PE=3 SV=2 <br/>
>sp|P37600|PHSA_SALTY Thiosulfate reductase OS=Salmonella typhimurium GN=phsA PE=3 SV=2 <br/>
-
MSISRRSFLQGVGIGCSACALGAFPPGALARNPIAGINGKTTLTPSLCEMCSFRCPIQAQ VVNNKTVFIQGNPSAPQQGTRICARGGSGVSLVNDPQRIVKPMKRTGPRGDGEWQVISWQ QAYQEIAAKMNAIKAQHGPESVAFSSKSGSLSSHLFHLATAFGSPNTFTHASTCPAGKAI AAKVMMGGDLAMDIANTRYLVSFGHNLYEGIEVADTHELMTAQEKGAKMVSFDPRLSIFS SKADEWHAIRPGGDLAVLLAMCHVMIDEQLYDASFVERYTSGFEQLAQAVKETTPEWAAA QADVPADVIVRVTRELAACAPHAIVSPGHRATFSQEEIDMRRMIFTLNVLLGNIEREGGL YQKKNASVYNKLAGEKVAPTLAKLNIKNMPKPTAQRIDLVAPQFKYIAAGGGVVQSIIDS ALTQKPYPIKAWIMSRHNPFQTVTCRSDLVKTVEQLDLVVSCDVYLSESAAYADYLLPEC TYLERDEEVSDMSGLHPAYALRQQVVEPIGEARPSWQIWKELGEQLGLGQYYPWQDMQTR QLYQLNGDHALAKELRQKGYLEWGVPLLLREPESVRQFTARYPGAIATDSDNTYGEQLRF KSPSGKIELYSATLEELLPGYGVPRVRDFALKKENELYFIQGKVAVHTNGATQYVPLLSE LMWDNAVWVHPQTAQEKGIKTGDEIWLENATGKEKGKALVTPGIRPDTLFVYMGFGAKAG AKTAATTHGIHCGNLLPHVTSPVSGTVVHTAGVTLSRA <br/>
+
MSISRRSFLQGVGIGCSACALGAFPPGALARNPIAGINGKTTLTPSLCEMCSFRCPIQAQ VVNNKTVFIQGNPSAPQQGTRICARGGSGVSLVNDPQRIVKPMKRTGPRGDGEWQVISWQ QAYQEIAAKMNAIKAQHGPESVAFSSKSGSLSSHLFHLATAFGSPNTFTHASTCPAGKAI AAKVMMGGDLAMDIANTRYLVSFGHNLYEGIEVADTHELMTAQEKGAKMVSFDPRLSIFS SKADEWHAIRPGGDLAVLLAMCHVMIDEQLYDASFVERYTSGFEQLAQAVKETTPEWAAA QADVPADVIVRVTRELAACAPHAIVSPGHRATFSQEEIDMRRMIFTLNVLLGNIEREGGL YQKKNASVYNKLAGEKVAPTLAKLNIKNMPKPTAQRIDLVAPQFKYIAAGGGVVQSIIDS ALTQKPYPIKAWIMSRHNPFQTVTCRSDLVKTVEQLDLVVSCDVYLSESAAYADYLLPEC TYLERDEEVSDMSGLHPAYALRQQVVEPIGEARPSWQIWKELGEQLGLGQYYPWQDMQTR QLYQLNGDHALAKELRQKGYLEWGVPLLLREPESVRQFTARYPGAIATDSDNTYGEQLRF KSPSGKIELYSATLEELLPGYGVPRVRDFALKKENELYFIQGKVAVHTNGATQYVPLLSE LMWDNAVWVHPQTAQEKGIKTGDEIWLENATGKEKGKALVTPGIRPDTLFVYMGFGAKAG
 +
AKTAATTHGIHCGNLLPHVTSPVSGTVVHTAGVTLSRA <br/>
>phsA <br/>
>phsA <br/>
-
atgagcattagtcgtcgttcttttttgcagggggtaggcatcggctgctctgcctgcgcgctgggcgcttttccgcccggggcgctggcgcgcaacccgattgccggcattaacggtaaaaccacgctaacgccaagcttgtgcgaaatgtgttcctttcgttgccctattcaggcgcaggtagtcaataacaagaccgtttttatccagggcaatccttctgcgccgcagcaggggacgcgcatatgcgccaggggcgggagcggcgtcagcctggtcaatgacccgcaacggattgtaaaacctatgaaacgcaccgggccacgcggcgacggtgaatggcaggtgattagctggcaacaggcttaccaggaaatcgcggcgaaaatgaatgccatcaaagcgcagcatggccccgagagcgtggctttctcttccaaatcgggctcgctctccagccatcttttccatctggctacggcctttggttcgcctaatacctttacgcacgcctcaacatgccctgccgggaaagccattgcggcaaaagtgatgatgggcggcgatctggcgatggatatcgctaacacgcgctatctggtttcgtttggccacaatttgtatgaagggattgaagttgccgatacccatgagttaatgaccgcgcaggagaagggggccaaaatggtgagcttcgatccgcgtttgtcgatattttccagcaaggcggatgagtggcacgctattcgtcccgggggggatttagcggttctgctggcgatgtgccacgtcatgattgatgaacagctctacgatgcgtcttttgttgagcgttataccagcggatttgaacagttagcacaggcggtaaaagagacgacgccggaatgggccgccgcgcaggccgatgtcccagccgacgttattgtccgggtgacacgcgaactggctgcctgcgcgcctcacgctattgtcagtcctggtcatcgcgcgacgttctcgcaggaagagatcgatatgcggcgtatgatttttacgcttaatgtgctgctcggtaatattgagcgcgaaggcgggctatatcagaaaaaaaacgcgtctgtttacaataaactggccggagaaaaggtcgcgccaacgctggcgaaactcaacattaaaaatatgccgaaaccgacggcgcaacgcatcgatttggtcgcaccgcagtttaaatatatcgccgctggcggcggcgtggtgcaaagcattattgactcggcgttaacccagaagccttacccgataaaggcgtggattatgtcgcggcataatcctttccagaccgtcacctgtcgttcggacctggtaaaaaccgttgagcaactggatctggtggtcagctgcgatgtctatttgagcgagagcgcggcatatgccgactatctgctgccggaatgcacctatctcgaacgggacgaagaggtatccgatatgtcgggactgcacccggcttacgctctgcgccaacaggtcgtagagccgattggcgaggcgcgtccgagttggcaaatctggaaagaacttggcgagcagttgggattagggcagtactatccgtggcaggatatgcagacgcgccaactctatcagttgaacggcgaccatgccttagcgaaggaactgcgacagaaagggtatctcgaatggggcgttccgctgctattacgcgaaccagaatccgttcgtcagtttacggcgcgttaccccggcgctatcgcgacggacagtgacaacacctatggcgaacagcttcgcttcaaatcgccctccggcaaaatcgaactttattccgcaaccctggaggaattgctccctggctacggcgttccgcgcgttcgtgactttgcgctgaaaaaagagaatgagctttacttcattcagggcaaggtggccgtgcataccaatggcgcgacgcagtacgtacctttactcagcgagctaatgtgggataacgcggtctgggttcatccgcaaacggcgcaagaaaaaggcattaagaccggcgatgagatctggctggaaaatgccacgggtaaagagaaaggtaaggcgctggtgacgcccggtatccgcccggacacgctttttgtctatatgggatttggcgctaaagctggggccaaaacggcggcgacgacacacggtatccactgcggaaatttactgccgcacgtgacgagtccggtatccggtacggtagtgcataccgcaggcgtgacgctgagccgggcatga <br/>
+
atgagcattagtcgtcgttcttttttgcagggggtaggcatcggctgctctgcctgcgcg ctgggcgcttttccgcccggggcgctggcgcgcaacccgattgccggcattaacggtaaa accacgctaacgccaagcttgtgcgaaatgtgttcctttcgttgccctattcaggcgcag gtagtcaataacaagaccgtttttatccagggcaatccttctgcgccgcagcaggggacg cgcatatgcgccaggggcgggagcggcgtcagcctggtcaatgacccgcaacggattgta aaacctatgaaacgcaccgggccacgcggcgacggtgaatggcaggtgattagctggcaa caggcttaccaggaaatcgcggcgaaaatgaatgccatcaaagcgcagcatggccccgag agcgtggctttctcttccaaatcgggctcgctctccagccatcttttccatctggctacg gcctttggttcgcctaatacctttacgcacgcctcaacatgccctgccgggaaagccatt gcggcaaaagtgatgatgggcggcgatctggcgatggatatcgctaacacgcgctatctg gtttcgtttggccacaatttgtatgaagggattgaagttgccgatacccatgagttaatg accgcgcaggagaagggggccaaaatggtgagcttcgatccgcgtttgtcgatattttcc agcaaggcggatgagtggcacgctattcgtcccgggggggatttagcggttctgctggcg atgtgccacgtcatgattgatgaacagctctacgatgcgtcttttgttgagcgttatacc agcggatttgaacagttagcacaggcggtaaaagagacgacgccggaatgggccgccgcg caggccgatgtcccagccgacgttattgtccgggtgacacgcgaactggctgcctgcgcg cctcacgctattgtcagtcctggtcatcgcgcgacgttctcgcaggaagagatcgatatg cggcgtatgatttttacgcttaatgtgctgctcggtaatattgagcgcgaaggcgggcta tatcagaaaaaaaacgcgtctgtttacaataaactggccggagaaaaggtcgcgccaacg ctggcgaaactcaacattaaaaatatgccgaaaccgacggcgcaacgcatcgatttggtc gcaccgcagtttaaatatatcgccgctggcggcggcgtggtgcaaagcattattgactcg gcgttaacccagaagccttacccgataaaggcgtggattatgtcgcggcataatcctttc cagaccgtcacctgtcgttcggacctggtaaaaaccgttgagcaactggatctggtggtc agctgcgatgtctatttgagcgagagcgcggcatatgccgactatctgctgccggaatgc acctatctcgaacgggacgaagaggtatccgatatgtcgggactgcacccggcttacgct ctgcgccaacaggtcgtagagccgattggcgaggcgcgtccgagttggcaaatctggaaa gaacttggcgagcagttgggattagggcagtactatccgtggcaggatatgcagacgcgc caactctatcagttgaacggcgaccatgccttagcgaaggaactgcgacagaaagggtat ctcgaatggggcgttccgctgctattacgcgaaccagaatccgttcgtcagtttacggcg cgttaccccggcgctatcgcgacggacagtgacaacacctatggcgaacagcttcgcttc aaatcgccctccggcaaaatcgaactttattccgcaaccctggaggaattgctccctggc tacggcgttccgcgcgttcgtgactttgcgctgaaaaaagagaatgagctttacttcatt cagggcaaggtggccgtgcataccaatggcgcgacgcagtacgtacctttactcagcgag ctaatgtgggataacgcggtctgggttcatccgcaaacggcgcaagaaaaaggcattaag accggcgatgagatctggctggaaaatgccacgggtaaagagaaaggtaaggcgctggtg acgcccggtatccgcccggacacgctttttgtctatatgggatttggcgctaaagctggg gccaaaacggcggcgacgacacacggtatccactgcggaaatttactgccgcacgtgacg agtccggtatccggtacggtagtgcataccgcaggcgtgacgctgagccgggcatga <br/>
 +
 
 +
phsB STM2064 <br/>
 +
>sp|P0A1I1|PHSB_SALTY Thiosulfate reductase electron transport protein phsB OS=Salmonella typhimurium GN=phsB PE=4 SV=1 <br/> MNHLTNQYVMLHDEKRCIGCQACTVACKVLNDVPEGFSRVQVQIRAPEQASNALTHFQFV RVSCQHCENAPCVSVCPTGASYRDENGIVQVDKSRCIGCDYCVAACPFHVRYLNPQTGVA DKCNFCADTRLAAGQSPACVSVCPTDALKFGRLDESEIQRWVGQKEVYRQQEARSGAVSL
 +
YRRKEVHQEGKA <br/>
 +
 
 +
>phsB <br/>
 +
Atgaatcatttaacgaatcagtacgtcatgctgcatgatgaaaaacgttgtatcggctgc
 +
caggcgtgtaccgttgcctgcaaagtgcttaatgacgtaccggagggatttagccgcgta
 +
caggtacaaattcgcgcccctgaacaggcatccaacgcactaacccattttcaatttgtc
 +
cgcgtctcctgtcagcactgtgaaaatgcgccatgtgtcagcgtttgtcctaccggagcg
 +
tcatatcgtgatgaaaacgggatcgtgcaggtggataaatcgcgctgtattggctgcgat
 +
tattgtgttgccgcgtgtcctttccatgtgcgctatttgaatccgcaaaccggtgtcgcc
 +
gacaagtgtaacttctgcgccgacacgcggttggcggctggccagtctccggcgtgcgta
 +
tccgtttgcccaacggacgcgctgaaattcggcagactggatgagagcgagatccagcgc
 +
tgggtcggtcagaaagaggtctatcgccagcaggaggcgcgtagcggcgcggtcagtctg
 +
taccgtcgtaaagaagtccatcaggagggtaaagcatga <br/>
 +
 
 +
phsC STM2063 <br/>
 +
>sp|P37602|PHSC_SALTY Thiosulfate reductase cytochrome B subunit OS=Salmonella typhimurium GN=phsC PE=4 SV=2 <br/>
 +
MNTIWGAELHYAPDYWPLWLIYAGVVVLLMLVGLVIHALLRRMLAPKTAGGEEHRDYLYS LAIRRWHWGNALLFVLLLLSGLFGHFSLGPVALMVQVHTWCGFALLAFWVGFVLINLTTG NGRHYRVNFSGLVTRCIRQTRFYLFGIMKGEAHPFVATEQNKFNPLQQLAYLAIMYALVP LLIITGLLCLYPQVAGLGPVMLVLHMALAIIGLLFICAHLYLCTLGDTPGQIFRSMVDGY
 +
HRHRTAPRGDKSAV <br/>
 +
 
 +
>phsC <br/>
 +
Atgaatactatctggggagcggaactacattatgcgccagattattggccgctgtggttaa
 +
tttacgcaggcgtcgtggtgctgctcatgcttgttgggctggttatccatgcgttattgcg
 +
ccggatgctggcgccaaaaacggcgggcggtgaagaacatcgtgactatctctactcgctg
 +
gcgattcgccgctggcattggggaaatgcgttactgtttgttttattactgttaagcggtt
 +
tatttggtcatttttctctcggccctgtagcgctaatggtacaagtgcatacctggtgtgg
 +
ttttgccttactggctttctgggtcgggtttgtgctgatcaacctcaccacaggtaacggg
 +
cgtcactatcgggtaaatttttccggactggtaacgcgctgcatacgccagacgcgttttt
 +
acctttttggcattatgaaaggggaagcgcatccgttcgtggcaacagagcagaataagtt
 +
caatccactgcaacaactggcatatctggcgattatgtacgcgctggtaccgctgttaatc
 +
atcaccggtttgctgtgtctctatccgcaggttgcgggtctgggccctgtgatgctggtgc
 +
tgcatatggcgcttgctatcatcggcttactgtttatttgcgcgcatctctatctgtgtac
 +
tcttggcgacacgccgggacaaattttccgtagcatggttgacggctatcatcgtcatcgt
 +
accgcgccgcgcggggataagtccgccgtctga <br/>
 +
 
<!------------- sequence data------------->
<!------------- sequence data------------->
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<br/>
<br/>
-
<i>This information is also recorded on page 11 of the hard copy lab notebook. </i>
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<!------------- Tuesday ------------->
<!------------- Tuesday ------------->
</div>
</div>

Revision as of 03:20, 27 October 2010

iGEM Yale

lab notebook: week 2 (6/14-6/20)

  • Monday 6/14--Redo of 6/11 assay of bacterial growth within a narrow copper(II) concentrations after analysis of data showed uniformly poor growth.
  • See more/less
  • Tuesday 6/15--Results and analysis of the 6/14 narrow concentration range growth assay as well as planning for the creation of a standard iGEM plasmid bearing the thiosulfate reductase operon (phsABC).
  • See more/less
  • Wednesday 6/16--checked on spotted cell survival assay, collected MOPS minimal media materials, started making component solutions
  • See more/less
  • Thursday 6/17--more minimal media work and meeting, started BL21 culture
  • See more/less
  • Friday 6/18--plasmid pSB74 arrived (!), already in some unspecified E coli, so plated them out. Transformed promoters (constitutive & IPTG inducible), terminator, and repressor (for inducible promoter) into BL21. Also ran BL21 copper growth assays for wide and narrow concentrations ranges. June the 19th--in which it is revealed that there was a transformation-fail Redid transformation, this time into LE392, set aside pSB74 containing cells in fridge
  • See more/less
  • Sunday 6/20--in evening inoculate 5 mL liquid cultures
  • See more/less