Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→vanillin synthesis)
(→Blue light receptor)
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| {{kulpart|BBa_E0240}}||pSB1A2||GFP + rbs + terminator||restricted with EcoRI and XbaI||||||july 29
| {{kulpart|BBa_E0240}}||pSB1A2||GFP + rbs + terminator||restricted with EcoRI and XbaI||||||july 29
 +
|-
 +
| {{kulpart|BBa_K145015}}||pSB1A2||GFP + LVA-tag + terminator||restricted with EcoRI and XbaI||||||july 29
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| {{kulpart|BBa_K238000}}||/||BLR promoter region (1:200 adn 1:500)||restricted with EcoRI||||||july 29
| {{kulpart|BBa_K238000}}||/||BLR promoter region (1:200 adn 1:500)||restricted with EcoRI||||||july 29
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| {{kulpart|BBa_K238000}}||/||PCR BLR promoter region (360bp)||4 microcentrifuge tubes||||||july 30
| {{kulpart|BBa_K238000}}||/||PCR BLR promoter region (360bp)||4 microcentrifuge tubes||||||july 30
|-
|-
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| {{iGempart|BBa_pSB3K3-P1010}}|| pSB3K3 ||kanamycine plasmide backbone|| taken from plates || 13μl ||||aug 11
+
| {{kulpart|BBa_pSB3K3-P1010}}|| pSB3K3 ||kanamycine plasmide backbone|| taken from plates || 13μl ||||aug 11
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|-
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| {{kulpart|BBa_K238013}}||/||Exact blue light promotor ||pcr product|| / || 123,3 ng/μl ||aug 13
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|-
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| {{kulpart|BBa_J23101}}||/||constitutive promotor||taken from plates || 13μl ||||aug 11
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|-
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| {{iGempart|BBa_J23101}}||/||constitutive promotor||taken from plates || 13μl ||||aug 11
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| {{kulpart|BBa_K238013}}||/||Exact blue light promotor ||cut with EcoRI and SpeI || 26μl || 27 ng/μl ||aug 12
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|-
-
| {{iGempart|BBa_K238013}}||/||Exact blue light promotor ||cut with EcoRI and SpeI || 26μl || 27 ng/μl ||aug 12
+
| {{kulpart|BBa_K238013+ BBa_E0240}}||/||LigA: Exact blue light promotor ligated with E0240 in pSB1A2 ||T4 DNA ligation || 18μl || ||aug 13
|-
|-
-
| {{iGempart|BBa_K238013+ BBa_E0240}}||/||Exact blue light promotor ligated with E0240 in pSB1A2 ||T4 DNA ligation || 18μl || ||aug 13
+
| {{kulpart|BBa_K238013+ BBa_K145015}}||/||Lig B: Exact blue light promotor ligated with K145015 in pSB1A2 ||T4 DNA ligation || 18μl || ||aug 14
|-
|-
-
| {{iGempart|BBa_K238013+ BBa_K145015}}||/||Exact blue light promotor ligated with K145015 in pSB1A2 ||T4 DNA ligation || 18μl || ||aug 14
+
| {{kulpart|BBa_K238013+ BBa_E02040}}||/||Lig A: Exact blue light promotor ligated with E02040 in pSB1A2 (1,3,5,7,9) ||cut with EcoRI and PstI || 5 x 18μl || ||aug 18
|}
|}

Revision as of 13:57, 19 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
pSB1A2GFP + LVA-tag + 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
pSB3K3 kanamycine plasmide backbone taken from plates 13μl aug 11
/Exact blue light promotor pcr product / 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 E0240 in pSB1A2 T4 DNA ligation 18μl aug 13
/Lig B: Exact blue light promotor ligated with K145015 in pSB1A2 T4 DNA ligation 18μl aug 14
/Lig A: Exact blue light promotor ligated with E02040 in pSB1A2 (1,3,5,7,9) cut with EcoRI and PstI 5 x 18μl aug 18

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
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
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
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
pSB1A2ech + rbspurified plasmid (ech A)36,8 µl88,6 ng/µlAugust 19
pSB1A2ech + rbspurified plasmid (ech B)48 µl96,7 ng/µlAugust 19
pSB1A2ech + rbspurified plasmid (ech C)48 µl105,6 ng/µlAugust 19
pSB1A2fcs + rbspurified plasmid (fcs A)30,3 µl84,8 ng/µlAugust 19
pSB1A2fcs + rbspurified plasmid (fcs B)48 µl113,4 ng/µlAugust 19
pSB1A2fcs + rbspurified plasmid (fcs C)48 µl90,3 ng/µlAugust 19
pSB1A2Sam5 + rbspurified plasmid (sam5 A)37,4 µl94,6 ng/µlAugust 19
pSB1A2Sam5 + rbspurified plasmid (sam5 B)48 µl125,0 ng/µlAugust 19
pSB1A2Sam5 + rbspurified plasmid (sam5 C)48 µl104,2 ng/µlAugust 19
pSB1A2Sam8 + rbspurified plasmid (sam8 A)21,9 µl58,2 ng/µlAugust 19
pSB1A2Sam8 + rbspurified plasmid (sam8 B)48 µl63,2 ng/µlAugust 19

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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