Team:KULeuven/Freezer

From 2009.igem.org

(Difference between revisions)
(→Blue light receptor)
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_E02040}} 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
Line 48: Line 48:
| {{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}}||pSB1A2||LigA (1): Exact blue light promotor ligated with E0240 in pSB1A2 ||Miniprep|| 45μl||110,6ng/μl||aug 31
+
| {{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}}||pSB1A2||LigA (2): Exact blue light promotor ligated with E0240 in pSB1A2 ||Miniprep|| 43μl||76,7ng/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}}||pSB1A2||LigA (3): Exact blue light promotor ligated with E0240 in pSB1A2 ||Miniprep|| 44μl||84,8ng/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}}||pSB1A2||LigA (4): Exact blue light promotor ligated with E0240 in pSB1A2 ||Miniprep|| 34μl||31,7ng/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_E0240}}||pSB1A2||RBS + GFP + terminator||Miniprep||45μl||104,3ng/μl||aug 31
Line 86: Line 86:
! class='top' | date
! class='top' | date
|-
|-
-
|{{kulpart|BBa_I742113}}||pSB1A2||ech + rbs||purified plasmid (oranje)||||108,8 ng/µl||August 4
+
|{{kulpart|BBa_I742113}}||{{kulpart|pSB1A2}}||ech + rbs||purified plasmid (oranje)||||108,8 ng/µl||August 4
|-
|-
-
|{{kulpart|BBa_K238007}}||pSB1A2||Sam5 + rbs, PstI site removed||purified plasmid (oranje)||||101,3 ng/µl||August 5
+
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||Sam5 + rbs, PstI site removed||purified plasmid (oranje)||||101,3 ng/µl||August 5
|-
|-
-
|{{kulpart|BBa_I742107}}||pSB1A2||COMT||purified plasmid (oranje)||||122,9 ng/µl||August 5
+
|{{kulpart|BBa_I742107}}||{{kulpart|pSB1A2}}||COMT||purified plasmid (oranje)||||122,9 ng/µl||August 5
|-
|-
-
|{{kulpart|BBa_I742115}}||pSB1A2||fcs + rbs||purified plasmid (fcs pl.)||29 µl||57,2 ng/µl||August 12
+
|{{kulpart|BBa_I742115}}||{{kulpart|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.)||31 µl||76,0 ng/µl||August 12
+
|{{kulpart|BBa_K238007}}||{{kulpart|pSB1A2}}||Sam5 + rbs||purified plasmid (sam5 pl.)||31 µl||76,0 ng/µl||August 12
|-
|-
-
|{{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 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}}||{{kulpart|pSB1A2}}||ech + rbs||purified plasmid (ech C)||48 µl||105,6 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}}||{{kulpart|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}}||{{kulpart|pSB1A2}}||fcs + rbs||purified plasmid (fcs C)||48 µl||90,3 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}}||{{kulpart|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}}||{{kulpart|pSB1A2}}||Sam5 + rbs||purified plasmid (sam5 C)||48 µl||104,2 ng/µl||August 19
|-
|-
-
|{{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 B)||40,1 µl||63,2 ng/µl||August 19
|-
|-
-
|||pSB1A2||sam8 + sam5||ligation mix (SAMS)||||||August 24
+
|||{{kulpart|pSB1A2}}||sam8 + sam5||ligation mix (SAMS)||||||August 24
|-
|-
-
|||pSB1A2||fcs + ech||ligation mix (EF)||||||August 24
+
|||{{kulpart|pSB1A2}}||fcs + ech||ligation mix (EF)||||||August 24
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF I)||48 µl||53,9 ng/µl||August 4
+
|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF I)||48 µl||53,9 ng/µl||August 4
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF II)||42 µl||39,4 ng/µl||August 4
+
|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF II)||42 µl||39,4 ng/µl||August 4
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF III)||39,9 µl||57,1 ng/µl||August 4
+
|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF III)||39,9 µl||57,1 ng/µl||August 4
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS I)||48 µl||76,8 ng/µl||August 4
+
|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS I)||48 µl||76,8 ng/µl||August 4
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS II)||35,3||83,4 ng/µl||August 4
+
|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS II)||35,3||83,4 ng/µl||August 4
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS III)||39,3 µl||61,5 ng/µl||August 4
+
|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS III)||39,3 µl||61,5 ng/µl||August 4
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS II), not purified from gel||5 µl|| ||August 31
+
|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS II), not purified from gel||5 µl|| ||August 31
|-
|-
-
|||pSB1A2||sam8 + sam5||purified ligation plasmid (SAMS I), purified from gel||25 µl|| ||August 31
+
|||{{kulpart|pSB1A2}}||sam8 + sam5||purified ligation plasmid (SAMS I), purified from gel||25 µl|| ||August 31
|-
|-
-
|||pSB1A2||fcs + ech||purified ligation plasmid (EF II), purified from gel||5 µl|| ||August 31
+
|||{{kulpart|pSB1A2}}||fcs + ech||purified ligation plasmid (EF II), purified from gel||5 µl|| ||August 31
|-
|-
|}
|}
Line 223: Line 223:
|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|J23110}}||{{kulpart|pSB1A2}}||medium constitutieve promoter||||||||july 23
|-
|-
|ampicilline||||||||||||
|ampicilline||||||||||||
|}
|}

Revision as of 12:55, 1 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

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
ech + rbspurified plasmid (oranje)108,8 ng/µlAugust 4
Sam5 + rbs, PstI site removedpurified plasmid (oranje)101,3 ng/µlAugust 5
COMTpurified plasmid (oranje)122,9 ng/µlAugust 5
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 + sam5ligation mix (SAMS)August 24
fcs + echligation mix (EF)August 24
fcs + echpurified ligation plasmid (EF I)48 µl53,9 ng/µlAugust 4
fcs + echpurified ligation plasmid (EF II)42 µl39,4 ng/µlAugust 4
fcs + echpurified ligation plasmid (EF III)39,9 µl57,1 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS I)48 µl76,8 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS II)35,383,4 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS III)39,3 µl61,5 ng/µlAugust 4
sam8 + sam5purified ligation plasmid (SAMS II), not purified from gel5 µl August 31
sam8 + sam5purified ligation plasmid (SAMS I), purified from gel25 µl August 31
fcs + echpurified ligation plasmid (EF II), purified from gel5 µl August 31

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
Retrieved from "http://2009.igem.org/Team:KULeuven/Freezer"