Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→vanillin receptor)
(→vanillin synthesis)
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|||pSB1A2||ech + fcs||ligation (lig.C)||||||August 11
|||pSB1A2||ech + fcs||ligation (lig.C)||||||August 11
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs pl.)||39 µl||57,2 ng/µl||August 12
+
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs pl.)||29 µl||57,2 ng/µl||August 12
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 pl.)||41 µl||76,0 ng/µl||August 12
+
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 pl.)||31 µl||76,0 ng/µl||August 12
|-
|-
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 pl.)||0 µl||45,2 ng/µl||August 12
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 pl.)||0 µl||45,2 ng/µl||August 12
|-
|-
-
|||||fcs + rbs||purified digest (F rd.)||28 µl||4,2 ng/µl||August 12
+
|||||fcs + rbs||purified digest (F rd.)||18 µl||4,2 ng/µl||August 12
|-
|-
-
|||pSB1A2||sam5 + rbs||purified digest (5 rd.)||28 µl||7,3 ng/µl||August 12
+
|||pSB1A2||sam5 + rbs||purified digest (5 rd.)||18 µl||7,3 ng/µl||August 12
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech pl.)||48 µl||72,7 ng/µl||August 13
+
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech pl.)||38 µl||72,7 ng/µl||August 13
|-
|-
-
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 pl.)||48 µl||66,2 ng/µl||August 13
+
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 pl.)||38 µl||66,2 ng/µl||August 13
|-
|-
|||pSB1A2||sam5 + sam8||cells with sam5+sam8 (elek. A)||600 µl||||August 13
|||pSB1A2||sam5 + sam8||cells with sam5+sam8 (elek. A)||600 µl||||August 13
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|||pSB1A2||ech + fcs||cells with ech+fcs (elek. C)||600 µl||||August 13
|||pSB1A2||ech + fcs||cells with ech+fcs (elek. C)||600 µl||||August 13
|-
|-
-
|||||sam8 + rbs||purified digest (sam8 rb.)||26 µl||9,4 ng/µl||August 14
+
|||||sam8 + rbs||purified digest (sam8 rd.)||16 µl||9,4 ng/µl||August 14
|-
|-
-
|||pSB1A2||ech + rbs||purified digest (ech rb.)||26 µl||13,6 ng/µl||August 14
+
|||pSB1A2||ech + rbs||purified digest (ech rd.)||16 µl||13,6 ng/µl||August 14
|-
|-
-
|||pSB1A2||sam8 + Sam5||ligation (SAMS)||28 µl|| ||August 14
+
|||pSB1A2||sam8 + Sam5||ligation (SAMS)||||||August 14
|-
|-
-
|||pSB1A2||ech + fcs||ligation (EF)||28 µl|| ||August 14
+
|||pSB1A2||ech + fcs||ligation (EF)||||||August 14
|}
|}

Revision as of 12:10, 18 August 2009

Contents

Blue light receptor

part plasmid description status μl left concentration date
MC4100 / strain of E.coli july 24
2171 / forward primer for 90µl 20μM july 24
2172 / reverse primer for 90µl 20μM july 24
2171 / forward primer for 100μM july 24
2172 / reverse primer for 100μM july 24
2260 / forward primer for 45µl 20μM aug 10
2261 / reverse primer for 45µl 20μM aug 10
2260 / forward primer for 100μM aug 10
2261 / reverse primer for 100μM aug 10
pSB1A2GFP + rbs + terminatorrestricted with EcoRI and XbaIjuly 29
/BLR promoter region (1:200 adn 1:500)restricted with EcoRIjuly 29
/PCR BLR promoter region (360bp)4 microcentrifuge tubesjuly 30
Template:IGempart pSB3K3 kanamycine plasmide backbone taken from plates 13μl aug 11
Template:IGempart/constitutive promotortaken from plates 13μl aug 11
Template:IGempart/Exact blue light promotor cut with EcoRI and SpeI 26μl 27 ng/μl aug 12
Template:IGempart/Exact blue light promotor ligated with E0240 in pSB1A2 T4 DNA ligation 18μl aug 13
Template:IGempart/Exact blue light promotor ligated with K145015 in pSB1A2 T4 DNA ligation 18μl aug 14

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
pSB1A2ech + rbspurified plasmid (oranje)108,8 ng/µlAugust 4
pSB1A2fcs + rbspurified plasmid (oranje)113,9 ng/µlAugust 4
pSB1A2Sam5 + rbsreceived from EdinburghAugust 3
pSB1A2Sam5 + rbs, PstI site removedpurified plasmid (oranje)101,3 ng/µlAugust 5
pSB1A2Sam8 + rbspurified plasmid (oranje)95,7 ng/µlAugust 5
pSB1A2Sam8 (b)purified plasmid (oranje)81,5 ng/µlAugust 5
pSB1A2COMTpurified plasmid (oranje)122,9 ng/µlAugust 5
pSB1A2COMT + rbsplasmid from registry0 µlAugust 6
pSB1A2ech + rbspurified digest (C2)16,9 ng/µlAugust 6
fcs + rbspurified digest (C1, FCS Rb)16,6 ng/µlAugust 6
pSB1A2sam5 + rbspurified digest (A2)3,1 ng/µlAugust 7
sam8 + rbspurified digest (A1)3,4 ng/µl August 7
pSB1A2sam5 + sam8ligation (lig. A)August 11
pSB1A2ech + fcsligation (lig.C)August 11
pSB1A2fcs + rbspurified plasmid (fcs pl.)29 µl57,2 ng/µlAugust 12
pSB1A2Sam5 + rbspurified plasmid (sam5 pl.)31 µl76,0 ng/µlAugust 12
pSB1A2Sam8 + rbspurified plasmid (sam8 pl.)0 µl45,2 ng/µlAugust 12
fcs + rbspurified digest (F rd.)18 µl4,2 ng/µlAugust 12
pSB1A2sam5 + rbspurified digest (5 rd.)18 µl7,3 ng/µlAugust 12
pSB1A2ech + rbspurified plasmid (ech pl.)38 µl72,7 ng/µlAugust 13
pSB1A2Sam8 + rbspurified plasmid (sam8 pl.)38 µl66,2 ng/µlAugust 13
pSB1A2sam5 + sam8cells with sam5+sam8 (elek. A)600 µlAugust 13
pSB1A2ech + fcscells with ech+fcs (elek. C)600 µlAugust 13
sam8 + rbspurified digest (sam8 rd.)16 µl9,4 ng/µlAugust 14
pSB1A2ech + rbspurified digest (ech rd.)16 µl13,6 ng/µlAugust 14
pSB1A2sam8 + Sam5ligation (SAMS)August 14
pSB1A2ech + fcsligation (EF)August 14

vanillin receptor

miscellaneous

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
part plasmid description status μl left concentration date
dNTP mixATP, GTP, TTP, CTP9410μMaug 10
pSB1A2medium constitutieve promoterjuly 23
ampicilline
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