Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→vanillin synthesis)
(→vanillin synthesis)
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|{{kulpart|BBa_I742106}}||{{kulpart|pSB1A2}}||Sam8 + rbs||purified plasmid (sam8 B)||40,1 µl||63,2 ng/µl||August 19
|{{kulpart|BBa_I742106}}||{{kulpart|pSB1A2}}||Sam8 + rbs||purified plasmid (sam8 B)||40,1 µl||63,2 ng/µl||August 19
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|||{{kulpart|pSB1A2}}||sam8 + sam5||ligation mix (SAMS)||||||August 24
 
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|||{{kulpart|pSB1A2}}||fcs + ech||ligation mix (EF)||||||August 24
 
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|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF I)||48 µl||53,9 ng/µl||August 4
 
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|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF II)||42 µl||39,4 ng/µl||August 4
 
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|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF III)||39,9 µl||57,1 ng/µl||August 4
 
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|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS I)||48 µl||76,8 ng/µl||August 4
 
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|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS II)||35,3||83,4 ng/µl||August 4
 
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|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS III)||39,3 µl||61,5 ng/µl||August 4
 
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|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS II), not purified from gel||5 µl|| ||August 31
 
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|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS I), purified from gel||25 µl|| ||August 31
 
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|||{{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')||16 µl ||35,4 ng/µl||September 3
|||{{kulpart|pSB1A2}}||sam8 + sam5||SAMS 1 (from 'mother plate')||16 µl ||35,4 ng/µl||September 3

Revision as of 14:09, 4 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
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 + sam5SAMS 1 (from 'mother plate')16 µl 35,4 ng/µlSeptember 3
sam8 + sam5SAMS 2 (from 'mother plate')19,9 µl49,3 ng/µl September 3
sam8 + sam5SAMS 3 (from 'mother plate')31,9 µl62,0 ng/µl September 3
sam8 + sam5SAMS 4 (from 'mother plate')33,6 µl78,3 ng/µl September 3
TerminatorTerminator, B0015 ±20 µl181,4 September 3
sam8SAM8 RD 3/928 µl 8,2 ng/µlSeptember 3
sam5SAM5 RD 3/9 28 µl1,4 ng/µl September 3
echECH RD 3/9 23 µl10,1 ng/µl September 3
fcsFCS RD 3/9 4 µl5,2 ng/µl 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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