Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→Blue light receptor)
(→vanillin synthesis)
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|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF II), purified from gel||5 µl|| ||August 31
|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF II), purified from gel||5 µl|| ||August 31
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|||{{kulpart|pSB1A2}}||sam8 + sam5||SAMS 1 (from 'mother plate')||35,3||35,4 ng/µl||September 3
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|||{{kulpart|pSB1A2}}||sam8 + sam5||SAMS 2 (from 'mother plate')||39,3 µl||49,3 ng/µl|| September 3
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|||{{kulpart|pSB1A2}}||sam8 + sam5||SAMS 3 (from 'mother plate')||5 µl||62,0 ng/µl|| September 3
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|||{{kulpart|pSB1A2}}||sam8 + sam5||SAMS 4 (from 'mother plate')||25 µl||78,3 ng/µl|| September 3
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|{{kulpart|BBa__B0015}}||{{kulpart|pSB1A2}}||Terminator||Terminator, B0015 || ±20 µl||181,4|| September 3
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Revision as of 14:05, 3 September 2009

Contents

Blue light receptor

part plasmid description status μl left concentration date
MC4100 / strain of E.coli july 24
2260 / forward primer for 42,5µl 20μM aug 10
2261 / reverse primer for 45µl 20μM aug 10
GFP + rbs + terminatorMiniprepped100μl85,6ng/μljuly 29
GFP + rbs + terminatorrestricted with EcoRI and XbaI 20μl july 29
GFP + LVA-tag + terminatorrestricted with EcoRI and XbaI30μl29,3 ng/μl aug 14
kanamycine plasmide backbone taken from plates 13μl aug 11
/Exact blue light promotor pcr product 20μl 123,3 ng/μl aug 13
/constitutive promotortaken from plates 13μl aug 11
/Exact blue light promotor cut with EcoRI and SpeI 26μl 27 ng/μl aug 12
+LigA: Exact blue light promotor ligated with in T4 DNA ligation 18μl aug 13
+Lig B: Exact blue light promotor ligated with in T4 DNA ligation 18μl aug 14
+/Lig A: Exact blue light promotor ligated with in (1,3,7,9) cut with EcoRI and PstI 4 x 18μl aug 24
+LigA: Exact blue light promotor ligated with in Miniprep (4x) 20μl aug 18
constitutive promotorminiprep10μl141,2ng/μlaug 14
(A)constitutive promotorrestriction digest with EcoRI en SpeI 20μl 20,8ng/μlaug 24
(B)constitutive promotorrestriction digest with EcoRI en SpeI 10μl 12,8ng/μlaug 24
+LigA (1): Exact blue light promotor ligated with in Miniprep 45μl110,6ng/μlaug 31
+LigA (2): Exact blue light promotor ligated with in Miniprep 43μl76,7ng/μlaug 31
+LigA (3): Exact blue light promotor ligated with in Miniprep 44μl84,8ng/μlaug 31
+LigA (4): Exact blue light promotor ligated with in Miniprep 34μl31,7ng/μlaug 31
pSB1A2RBS + GFP + terminatorMiniprep45μl104,3ng/μlaug 31
(A)pSB1A2 promotorMiniprep48μl238,9ng/μlsept 2
(B)pSB1A2 promotorMiniprep48μl248,8ng/μlsept 2

Key/antikey/lock

part plasmid description status μl left concentration date
/GFP with LVA tagpurifiedjuly 28
/tetR promoter purifiedjuly 28

vanillin synthesis

part plasmid description status μl left concentration date
ech + rbspurified plasmid (oranje)108,8 ng/µlAugust 4
Sam5 + rbs, PstI site removedpurified plasmid (oranje)101,3 ng/µlAugust 5
COMTpurified plasmid (oranje)122,9 ng/µlAugust 5
fcs + rbspurified plasmid (fcs pl.)29 µl57,2 ng/µlAugust 12
Sam5 + rbspurified plasmid (sam5 pl.)31 µl76,0 ng/µlAugust 12
ech + rbspurified plasmid (ech B)42,8 µl96,7 ng/µlAugust 19
ech + rbspurified plasmid (ech C)48 µl105,6 ng/µlAugust 19
fcs + rbspurified plasmid (fcs B)43,6 µl113,4 ng/µlAugust 19
fcs + rbspurified plasmid (fcs C)48 µl90,3 ng/µlAugust 19
Sam5 + rbspurified plasmid (sam5 B)44 µl125,0 ng/µlAugust 19
Sam5 + rbspurified plasmid (sam5 C)48 µl104,2 ng/µlAugust 19
Sam8 + rbspurified plasmid (sam8 B)40,1 µl63,2 ng/µlAugust 19
sam8 + sam5ligation mix (SAMS)August 24
fcs + echligation mix (EF)August 24
fcs + echpurified ligation plasmid (EF I)48 µl53,9 ng/µlAugust 4
fcs + echpurified ligation plasmid (EF II)42 µl39,4 ng/µlAugust 4
fcs + echpurified ligation plasmid (EF III)39,9 µl57,1 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS I)48 µl76,8 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS II)35,383,4 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS III)39,3 µl61,5 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS II), not purified from gel5 µl August 31
sam8 + sam5purified ligation plasmid (SAMS I), purified from gel25 µl August 31
fcs + echpurified ligation plasmid (EF II), purified from gel5 µl August 31
sam8 + sam5SAMS 1 (from 'mother plate')35,335,4 ng/µlSeptember 3
sam8 + sam5SAMS 2 (from 'mother plate')39,3 µl49,3 ng/µl September 3
sam8 + sam5SAMS 3 (from 'mother plate')5 µl62,0 ng/µl September 3
sam8 + sam5SAMS 4 (from 'mother plate')25 µl78,3 ng/µl September 3
TerminatorTerminator, B0015 ±20 µl181,4 September 3

vanillin receptor

part plasmid description status μl left concentration date
2235 / forward primer for 20μM August 3
2238 / reverse primer for 20μM August 3
2235 / forward primer for 100μM August 3
2238 / reverse primer for 100μM August 3
2242 / forward primer for 20μM August 3
2243 / reverse primer for 20μM August 3
2242 / forward primer for 100μM August 3
2243 / reverse primer for 100μM August 3
2244 / forward primer for 20μM August 3
2245 / reverse primer for 20μM August 3
2244 / forward primer for 100μM August 3
2245 / reverse primer for 100μM August 3
2246 / forward primer for 20μM August 3
2247 / reverse primer for 20μM August 3
2246 / forward primer for 100μM August 3
2247 / reverse primer for 100μM August 3
2288 / forward primer for mutagenesis for 20μM August 8
2289 / reverse primer for mutagenesis for 20μM August 8
2288 / forward primer for mutagenesis for 100μM August 8
2289 / reverse primer for mutagenesis for 100μM August 8
2290 / forward primer for mutagenesis for 20μM August 8
2291 / reverse primer for mutagenesis for 20μM August 8
2290 / forward primer for mutagenesis for 100μM August 8
2291 / reverse primer for mutagenesis for 100μM August 8
2292 / forward primer for mutagenesis for 20μM August 8
2293 / reverse primer for mutagenesis for 20μM August 8
2292 / forward primer for mutagenesis for 100μM August 8
2293 / reverse primer for mutagenesis for 100μM August 8
2294 / forward primer for mutagenesis for 20μM August 8
2295 / reverse primer for mutagenesis for 20μM August 8
2294 / forward primer for mutagenesis for 100μM August 8
2295 / reverse primer for mutagenesis for 100μM August 8

miscellaneous

part plasmid description status μl left concentration date
dNTP mixATP, GTP, TTP, CTP9410μMaug 10
medium constitutieve promoterjuly 23
ampicilline
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