Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→Blue light receptor)
(→vanillin synthesis)
 
(20 intermediate revisions not shown)
Line 20: Line 20:
| 2261 || / || reverse primer for {{kulpart|BBa_K238013}} || || 45µl || 20μM || aug 10
| 2261 || / || reverse primer for {{kulpart|BBa_K238013}} || || 45µl || 20μM || aug 10
|-
|-
-
| {{kulpart|BBa_E0240}}||pSB1A2||GFP + rbs + terminator||Miniprepped||100μl||85,6ng/μl||july 29
+
| {{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||GFP + rbs + terminator||Miniprepped||100μl||85,6ng/μl||july 29
|-
|-
-
| {{kulpart|BBa_E0240}}||pSB1A2||GFP + rbs + terminator||restricted with EcoRI and XbaI|| 20μl ||||july 29
+
| {{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||GFP + rbs + terminator||restricted with EcoRI and XbaI|| 20μl ||||july 29
|-
|-
-
| {{kulpart|BBa_K145015}}||pSB1A2||GFP + LVA-tag + terminator||restricted with EcoRI and XbaI||30μl||29,3 ng/μl|| aug 14
+
| {{kulpart|BBa_K145015}}||{{kulpart|pSB1A2}}||GFP + LVA-tag + terminator||restricted with EcoRI and XbaI||30μl||29,3 ng/μl|| aug 14
|-
|-
-
| {{kulpart|pSB3K3}}|| pSB3K3 ||kanamycine plasmide backbone|| taken from plates || 13μl ||||aug 11
+
| {{kulpart|pSB3K3}}|| {{kulpart|pSB3K3}} ||kanamycine plasmide backbone|| taken from plates || 13μl ||||aug 11
|-
|-
| {{kulpart|BBa_K238013}}||/||Exact blue light promotor ||pcr product|| 20μl || 123,3 ng/μl ||aug 13
| {{kulpart|BBa_K238013}}||/||Exact blue light promotor ||pcr product|| 20μl || 123,3 ng/μl ||aug 13
Line 34: Line 34:
| {{kulpart|BBa_K238013}}||/||Exact blue light promotor ||cut with EcoRI and SpeI || 26μl || 27 ng/μl ||aug 12
| {{kulpart|BBa_K238013}}||/||Exact blue light promotor ||cut with EcoRI and SpeI || 26μl || 27 ng/μl ||aug 12
|-
|-
-
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||pSB1A3||LigA: Exact blue light promotor ligated with E0240 in pSB1A2 ||T4 DNA ligation || 18μl || ||aug 13
+
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||{{kulpart|pSB1A3}}||LigA: Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} ||T4 DNA ligation || 18μl || ||aug 13
|-
|-
-
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_K145015}}||pSB1A3||Lig B: Exact blue light promotor ligated with K145015 in pSB1A2 ||T4 DNA ligation || 18μl || ||aug 14
+
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_K145015}}||{{kulpart|pSB1A3}}||Lig B: Exact blue light promotor ligated with {{kulpart|BBa_K145015}} in {{kulpart|pSB1A2}} ||T4 DNA ligation || 18μl || ||aug 14
|-
|-
-
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||/||Lig A: Exact blue light promotor ligated with E02040 in pSB1A2 (1,3,7,9) ||cut with EcoRI and PstI || 4 x 18μl || ||aug 24
+
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||/||Lig A: Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} (1,3,7,9) ||cut with EcoRI and PstI || 4 x 18μl || ||aug 24
|-
|-
-
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||pSB1A2||LigA: Exact blue light promotor ligated with E0240 in pSB1A2 ||Miniprep (4x)|| 20μl|| ||aug 18
+
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||LigA: Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} ||Miniprep (4x)|| 20μl|| ||aug 18
|-
|-
-
| {{kulpart|BBa_J23101}}||pSB1A3||constitutive promotor||miniprep||10μl||141,2ng/μl||aug 14
+
| {{kulpart|BBa_J23101}}||{{kulpart|pSB1A3}}||constitutive promotor||miniprep||10μl||141,2ng/μl||aug 14
|-
|-
| {{kulpart|BBa_J23101}}(A)||||constitutive promotor||restriction digest with EcoRI en SpeI|| 20μl ||20,8ng/μl||aug 24
| {{kulpart|BBa_J23101}}(A)||||constitutive promotor||restriction digest with EcoRI en SpeI|| 20μl ||20,8ng/μl||aug 24
|-
|-
| {{kulpart|BBa_J23101}}(B)||||constitutive promotor||restriction digest with EcoRI en SpeI|| 10μl ||12,8ng/μl||aug 24
| {{kulpart|BBa_J23101}}(B)||||constitutive promotor||restriction digest with EcoRI en SpeI|| 10μl ||12,8ng/μl||aug 24
 +
|-
 +
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||LigA (1): Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} ||Miniprep|| 45μl||110,6ng/μl||aug 31
 +
|-
 +
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||LigA (2): Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} ||Miniprep|| 43μl||76,7ng/μl||aug 31
 +
|-
 +
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||LigA (3): Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} ||Miniprep|| 44μl||84,8ng/μl||aug 31
 +
|-
 +
| {{kulpart|BBa_K238013}}+{{kulpart|BBa_E0240}}||{{kulpart|pSB1A2}}||LigA (4): Exact blue light promotor ligated with {{kulpart|BBa_E0240}} in {{kulpart|pSB1A2}} ||Miniprep|| 34μl||31,7ng/μl||aug 31
 +
|-
 +
| {{kulpart|BBa_E0240}}||pSB1A2||RBS + GFP + terminator||Miniprep||45μl||104,3ng/μl||aug 31
 +
|-
 +
| {{kulpart|BBa_J23101}} (A)||pSB1A2|| promotor||Miniprep||48μl||238,9ng/μl||sept 2
 +
|-
 +
| {{kulpart|BBa_J23101}} (B)||pSB1A2|| promotor||Miniprep||48μl||248,8ng/μl||sept 2
|}
|}
Line 76: Line 90:
! class='top' | date
! class='top' | date
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (oranje)||||108,8 ng/µl||August 4
+
|{{kulpart|BBa_I742115}}||{{kulpart|pSB1A2}}||fcs + rbs||purified plasmid (FCS 1)||13 µl||32.1 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs, PstI site removed||purified plasmid (oranje)||||101,3 ng/µl||August 5
+
|{{kulpart|BBa_I742115}}||{{kulpart|pSB1A2}}||fcs + rbs||purified plasmid (FCS 2)||28 µl||67.2 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742107}}||pSB1A2||COMT||purified plasmid (oranje)||||122,9 ng/µl||August 5
+
|{{kulpart|BBa_I742115}}||{{kulpart|pSB1A2}}||fcs + rbs||purified plasmid (FCS 3)||28 µl||82.4 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs pl.)||29 µl||57,2 ng/µl||August 12
+
|{{kulpart|BBa_I742113}}||{{kulpart|pSB1A2}}||ech + rbs||purified plasmid (ECH 1)||15 µl||37.2 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 pl.)||31 µl||76,0 ng/µl||August 12
+
|{{kulpart|BBa_I742113}}||{{kulpart|pSB1A2}}||ech + rbs||purified plasmid (ECH 2)||28 µl||26.9 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech B)||42,8 µl||96,7 ng/µl||August 19
+
|{{kulpart|BBa_I742113}}||{{kulpart|pSB1A2}}||ech + rbs||purified plasmid (ECH 3)||28 µl||28.0 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (ech C)||48 µl||105,6 ng/µl||August 19
+
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||Sam5 + rbs||purified plasmid (SAM5 1)||19.6 µl||59.9 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs B)||43,6 µl||113,4 ng/µl||August 19
+
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||Sam5 + rbs||purified plasmid (SAM5 2)||28 µl||52.8 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs C)||48 µl||90,3 ng/µl||August 19
+
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||Sam5 + rbs||purified plasmid (SAM5 3)||13 µl||67.3 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 B)||44 µl||125,0 ng/µl||August 19
+
|{{kulpart|BBa_I742106}}||{{kulpart|pSB1A2}}||Sam8 + rbs||purified plasmid (SAM8 1)||12 µl||31.0 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs||purified plasmid (sam5 C)||48 µl||104,2 ng/µl||August 19
+
|{{kulpart|BBa_I742106}}||{{kulpart|pSB1A2}}||Sam8 + rbs||purified plasmid (SAM8 2)||8 µl||47.2 ng/µl||September 1
|-
|-
-
|{{kulpart|BBa_I742106}}||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 3)||28 µl||42.9 ng/µl||September 1
|-
|-
-
|||pSB1A2||sam8 + sam5||ligation mix (SAMS)||||||August 24
+
|{{kulpart|BBa__B0015}}||{{kulpart|pSB1AK3}}||Terminator||Terminator, B0015 || ±20 µl||181,4|| September 3
|-
|-
-
|||pSB1A2||fcs + ech||ligation mix (EF)||||||August 24
+
|{{kulpart|BBa_I742106}}||{{kulpart|pSB1A2}}||sam8||SAM8 RD 3/9||28 µl ||8,2 ng/µl||September 3
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF I)||48 µl||53,9 ng/µl||August 4
+
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||sam5||SAM5 RD 3/9 ||28 µl||1,4 ng/µl|| September 3
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF II)||42 µl||39,4 ng/µl||August 4
+
|{{kulpart|BBa_I742113}}||{{kulpart|pSB1A2}}||ech||ECH RD 3/9 ||23 µl||10,1 ng/µl|| September 3
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF III)||39,9 µl||57,1 ng/µl||August 4
+
|{{kulpart|BBa_I742115}}||{{kulpart|pSB1A2}}||fcs||FCS RD 3/9 ||4 µl||5,2 ng/µl|| September 3
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS I)||48 µl||76,8 ng/µl||August 4
+
|{{kulpart|BBa_I742106}}||||sam8||SAM8 PCR||µl ||361.6 ng/µl||September 3
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS II)||35,3||83,4 ng/µl||August 4
+
|{{kulpart|BBa_K238007}}||||sam5||SAM5 PCR||µl||323.0 ng/µl|| September 3
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS III)||39,3 µl||61,5 ng/µl||August 4
+
|{{kulpart|BBa_I742113}}||||ech||ECH PCR||µl||236.5 ng/µl|| September 3
|-
|-
 +
|{{kulpart|BBa_I742115}}||||fcs||FCS PCR||µl||321.4 ng/µl|| September 3
 +
|-
 +
|||{{kulpart|pSB1A2}}||ech + fcs||Ligatie zonder terminator(EFS)|| µl||ng/µl|| September 3
 +
|-
 +
|||{{kulpart|pSB1A2}}||ech + fcs||Ligatie met terminator(EFT) || µl||ng/µl|| September 3
 +
|-
 +
|||{{kulpart|pSB1A2}}||sam5 + sam8||Ligatie zonder terminator(58S)|| µl||ng/µl|| September 4
 +
|-
 +
|||{{kulpart|pSB1A2}}||sam5 + sam8||Ligatie met terminator(58T)|| µl||ng/µl|| September 4
 +
|-
 +
|||{{kulpart|pSB1A2}}||ech + fcs||Miniprepped Ligatie Jochem (EF1)||48 µl|| 115.5 ng/µl|| September 7
 +
|-
 +
|||{{kulpart|pSB1A2}}||ech + fcs||Miniprepped Ligatie Jochem (EF3)||48 µl|| 95.1 ng/µl|| September 7
 +
|-
 +
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||sam5||SAM5 purified||23 µl||10.0 ng/µl|| September 7
 +
|-
 +
|||{{kulpart|pSB1A2}}||ech + fcs||Restrictie Ligatie EF - Jochem (EFX 3)|| 22 µl|| 4,3 ng/µl|| September 9
 +
|-
 +
|{{kulpart|BBa__B0015}}||{{kulpart|pSB1AK3}}||TER||Restrictie TER - Jochem (TER RD)|| 24 µl|| 14,2 ng/µl|| September 10
 +
|-
 +
|||{{kulpart|pSB1A2}}||Sam5+Sam8||Restrictie Sam5/Sam8 - Jochem E/S (SAMS1)|| 28 µl|| X ng/µl|| September 10
 +
|-
 +
|}
|}
Line 207: Line 244:
|dNTP mix||||ATP, GTP, TTP, CTP||||94||10μM||aug 10
|dNTP mix||||ATP, GTP, TTP, CTP||||94||10μM||aug 10
|-
|-
-
|{{kulpart|J23110}}||pSB1A2||medium constitutieve promoter||||||||july 23
+
|{{kulpart|BBa_J23110}}||{{kulpart|pSB1A2}}||medium constitutieve promoter||||||||july 23
|-
|-
|ampicilline||||||||||||
|ampicilline||||||||||||
|}
|}

Latest revision as of 11:20, 14 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 1)13 µl32.1 ng/µlSeptember 1
fcs + rbspurified plasmid (FCS 2)28 µl67.2 ng/µlSeptember 1
fcs + rbspurified plasmid (FCS 3)28 µl82.4 ng/µlSeptember 1
ech + rbspurified plasmid (ECH 1)15 µl37.2 ng/µlSeptember 1
ech + rbspurified plasmid (ECH 2)28 µl26.9 ng/µlSeptember 1
ech + rbspurified plasmid (ECH 3)28 µl28.0 ng/µlSeptember 1
Sam5 + rbspurified plasmid (SAM5 1)19.6 µl59.9 ng/µlSeptember 1
Sam5 + rbspurified plasmid (SAM5 2)28 µl52.8 ng/µlSeptember 1
Sam5 + rbspurified plasmid (SAM5 3)13 µl67.3 ng/µlSeptember 1
Sam8 + rbspurified plasmid (SAM8 1)12 µl31.0 ng/µlSeptember 1
Sam8 + rbspurified plasmid (SAM8 2)8 µl47.2 ng/µlSeptember 1
Sam8 + rbspurified plasmid (SAM8 3)28 µl42.9 ng/µlSeptember 1
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
sam8SAM8 PCRµl 361.6 ng/µlSeptember 3
sam5SAM5 PCRµl323.0 ng/µl September 3
echECH PCRµl236.5 ng/µl September 3
fcsFCS PCRµl321.4 ng/µl September 3
ech + fcsLigatie zonder terminator(EFS) µlng/µl September 3
ech + fcsLigatie met terminator(EFT) µlng/µl September 3
sam5 + sam8Ligatie zonder terminator(58S) µlng/µl September 4
sam5 + sam8Ligatie met terminator(58T) µlng/µl September 4
ech + fcsMiniprepped Ligatie Jochem (EF1)48 µl 115.5 ng/µl September 7
ech + fcsMiniprepped Ligatie Jochem (EF3)48 µl 95.1 ng/µl September 7
sam5SAM5 purified23 µl10.0 ng/µl September 7
ech + fcsRestrictie Ligatie EF - Jochem (EFX 3) 22 µl 4,3 ng/µl September 9
TERRestrictie TER - Jochem (TER RD) 24 µl 14,2 ng/µl September 10
Sam5+Sam8Restrictie Sam5/Sam8 - Jochem E/S (SAMS1) 28 µl X ng/µl September 10

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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