Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→vanillin synthesis)
Line 128: Line 128:
|||pSB1A2||ech + fcs||ligation (EF)||||||August 14
|||pSB1A2||ech + fcs||ligation (EF)||||||August 14
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech A)||36,8 µl||88,6 ng/µl||August 19
+
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech A)||0 µl||88,6 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech B)||48 µl||96,7 ng/µl||August 19
+
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech B)||42,8 µl||96,7 ng/µl||August 19
|-
|-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech C)||48 µl||105,6 ng/µl||August 19
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech C)||48 µl||105,6 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs A)||30,3 µl||84,8 ng/µl||August 19
+
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs A)||0 µl||84,8 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs B)||48 µl||113,4 ng/µl||August 19
+
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs B)||43,6 µl||113,4 ng/µl||August 19
|-
|-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs C)||48 µl||90,3 ng/µl||August 19
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs C)||48 µl||90,3 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 A)||37,4 µl||94,6 ng/µl||August 19
+
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 A)||0 µl||94,6 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 B)||48 µl||125,0 ng/µl||August 19
+
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 B)||44 µl||125,0 ng/µl||August 19
|-
|-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 C)||48 µl||104,2 ng/µl||August 19
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 C)||48 µl||104,2 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 A)||21,9 µl||58,2 ng/µl||August 19
+
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 A)||0 µl||58,2 ng/µl||August 19
|-
|-
-
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 B)||48 µl||63,2 ng/µl||August 19
+
|{{kulpart|BBa_I742106}}||pSB1A2||Sam8 + rbs||purified plasmid (sam8 B)||40,1 µl||63,2 ng/µl||August 19
 +
|-
 +
|||pSB1A2||fcs + ech||purified ligated plasmid (EF A)||48 µl||74,9 ng/µl||August 20
 +
|-
 +
|||pSB1A2||fcs + ech||purified ligated plasmid (EF B)||48 µl||63,1 ng/µl||August 20
 +
|-
 +
|||pSB1A2||fcs + ech||purified ligated plasmid (EF C)||38,7 µl||53,9 ng/µl||August 20
|}
|}

Revision as of 13:47, 20 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 EdinburghLost in transitAugust 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)0 µl88,6 ng/µlAugust 19
pSB1A2ech + rbspurified plasmid (ech B)42,8 µl96,7 ng/µlAugust 19
pSB1A2ech + rbspurified plasmid (ech C)48 µl105,6 ng/µlAugust 19
pSB1A2fcs + rbspurified plasmid (fcs A)0 µl84,8 ng/µlAugust 19
pSB1A2fcs + rbspurified plasmid (fcs B)43,6 µl113,4 ng/µlAugust 19
pSB1A2fcs + rbspurified plasmid (fcs C)48 µl90,3 ng/µlAugust 19
pSB1A2Sam5 + rbspurified plasmid (sam5 A)0 µl94,6 ng/µlAugust 19
pSB1A2Sam5 + rbspurified plasmid (sam5 B)44 µl125,0 ng/µlAugust 19
pSB1A2Sam5 + rbspurified plasmid (sam5 C)48 µl104,2 ng/µlAugust 19
pSB1A2Sam8 + rbspurified plasmid (sam8 A)0 µl58,2 ng/µlAugust 19
pSB1A2Sam8 + rbspurified plasmid (sam8 B)40,1 µl63,2 ng/µlAugust 19
pSB1A2fcs + echpurified ligated plasmid (EF A)48 µl74,9 ng/µlAugust 20
pSB1A2fcs + echpurified ligated plasmid (EF B)48 µl63,1 ng/µlAugust 20
pSB1A2fcs + echpurified ligated plasmid (EF C)38,7 µl53,9 ng/µlAugust 20

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
pSB1A2medium constitutieve promoterjuly 23
ampicilline
Retrieved from "http://2009.igem.org/Team:KULeuven/Freezer"