Paris/August 16

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[[Paris/August 15|yesterday]] -- [[Paris/August 17|tomorrow]]
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==Colony PCRs==
==Colony PCRs==
The cloning finally seem to have worked, so we are doing colony PCRs to check if there is an insert and if it's got the right size.
The cloning finally seem to have worked, so we are doing colony PCRs to check if there is an insert and if it's got the right size.
 +
 +
 +
== Digestion reactions ==
 +
 +
The following digestion reactions (ON) were performed:
 +
 +
<br>
 +
<font color="#FF99FF" size="4">PURPLE = Vector </font><br>
 +
<font color="#ccffcc" size="4">GREEN = Insert </font><br>
 +
{|
 +
|- style="background: #ccccff;"
 +
! colspan="5" style="background: #ccccff;" | Digestion Products
 +
|- style="background: #ccccff; text-align: center;"
 +
|width=5%| Number
 +
|width=24%| Product Name
 +
|width=20%| Matrix Name
 +
|width=5%| Enzyme 1
 +
|width=5%| Enzyme 2
 +
|width=5%| Size
 +
|width=34%| Description
 +
|width=1%|
 +
|width=1%|
 +
|width=5%| Date
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D37
 +
|lox71-gfp(tripart) BI
 +
|L12.2 miniprep products (from [[Paris/July 30| 30/07]] ligation reaction)
 +
|XbaI
 +
|PstI
 +
|
 +
|lox71_RBS-gfp-double terminator extracted as a backward insert
 +
|
 +
|
 +
|Aug. 16
 +
|}
 +
 +
{|
 +
|- style="background: #ccccff;"
 +
! colspan="5" style="background: #ccccff;" | Digestion Products
 +
|- style="background: #ccccff; text-align: center;"
 +
|width=5%| Number
 +
|width=20%| Product Name
 +
|width=24%| Matrix Name
 +
|width=5%| Enzyme 1
 +
|width=5%| Enzyme 2
 +
|width=5%| Size
 +
|width=38%| Description
 +
|width=1%|
 +
|width=1%|
 +
|width=1%|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D44
 +
|<bbpart>B0015</bbpart> terminator BI
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> (MP3.3 Aug.14)
 +
|XbaI
 +
|Pst1
 +
|
 +
|terminator B0015 extracted for use as a BI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D45
 +
|<bbpart>B0015</bbpart> terminator FI
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> (MP3.4 Aug.14)
 +
|EcoRI
 +
|SpeI
 +
|
 +
|terminator B0015 extracted for use as a FI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D46
 +
|<bbpart>B0015</bbpart> BV
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> (MP3.3 Aug.14)
 +
|SpeI
 +
|Pst1
 +
|
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> plasmid extracted for use as a BV
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D47
 +
|<bbpart>pSB1A2</bbpart> XP dig.
 +
|<bbpart>pSB1A2</bbpart> carrying Cre ORF (MP9.1)
 +
|XbaI
 +
|PstI
 +
|
 +
|<bbpart>pSB1A2</bbpart> open vector ready for cloning of BI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D48
 +
|<bbpart>pSB1A2</bbpart> ES dig.
 +
|<bbpart>pSB1A2</bbpart> carrying Cre ORF (MP9.2)
 +
|EcoRI
 +
|SpeI
 +
|
 +
|<bbpart>pSB1A2</bbpart> open vector ready for cloning of FI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D49
 +
|lox71-gfptripart BI
 +
|lox71-gfptripart in <bbpart>pSB1A2</bbpart> (L12.2)
 +
|XbaI
 +
|Pst1
 +
|
 +
|lox71-gfptripart extracted for use as a BI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D50
 +
|<bbpart>BBa_J61002</bbpart> pTet SP dig.
 +
|<bbpart>BBa_J61002</bbpart> (MP13.1 [[Paris/August 14| Aug. 14]])
 +
|SpeI
 +
|PstI
 +
|
 +
|<bbpart>BBa_J61002</bbpart> pTet promoter vector ready for insertion of CDS as a BI
 +
|
 +
|
 +
|
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|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D51
 +
|RBS-dapAcoli BBinsert
 +
|pTOPO of RBS-dapAcoli (MPT4.3)
 +
|EcoRI
 +
|Pst1
 +
|
 +
|RBS-dapAcoli extracted for cloning as a biobrick
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D52
 +
|RBS-dapAcoli BI
 +
|pTOPO of RBS-dapAcoli (MPT4.3)
 +
|XbaI
 +
|Pst1
 +
|
 +
|RBS-dapAcoli extracted for use as a BI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D53
 +
|RBS-dapAsubtilis BBinsert
 +
|pTOPO of RBS-dapAsubtilis (MPT5.1)
 +
|EcoRI
 +
|Pst1
 +
|
 +
|RBS-dapAsubtilis extracted for cloning as a biobrick
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D54
 +
|RBS-dapAsubtilis BI
 +
|pTOPO of RBS-dapAsubtilis (MPT5.1)
 +
|XbaI
 +
|Pst1
 +
|
 +
|RBS-dapAsubtilis extracted for use as a BI
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D55
 +
|RBS-dapAsubtilis FI
 +
|pTOPO of RBS-dapAsubtilis (MPT5.2)
 +
|EcoRI
 +
|SpeI
 +
|
 +
|RBS-dapAsubtilis extracted for use as a FI
 +
|
 +
|
 +
|
 +
|}
 +
 +
 +
 +
 +
I was unable to perform the following digestion reactions (because their was no more MP1 or MP2 product left):
 +
 +
<br>
 +
<font color="#FF99FF" size="4">PURPLE = Vector </font><br>
 +
<font color="#ccffcc" size="4">GREEN = Insert </font><br>
 +
{|
 +
|- style="background: #ccccff;"
 +
! colspan="5" style="background: #ccccff;" | Digestion Products
 +
|- style="background: #ccccff; text-align: center;"
 +
|width=5%| Number
 +
|width=24%| Product Name
 +
|width=20%| Matrix Name
 +
|width=5%| Enzyme 1
 +
|width=5%| Enzyme 2
 +
|width=5%| Size
 +
|width=34%| Description
 +
|width=1%|
 +
|width=1%|
 +
|width=5%| Date
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D1
 +
|<bbpart>pJ23100</bbpart>
 +
|<bbpart>pJ23100</bbpart>
 +
|Spe1
 +
|Pst1
 +
|2096
 +
|Vector with the J23100 promoter ready for insertion of a RBS/CDS insert
 +
|
 +
|
 +
|
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D3
 +
|<bbpart>pJ23107</bbpart>
 +
|<bbpart>pJ23107</bbpart>
 +
|Spe1
 +
|Pst1
 +
|2096
 +
|Vector with the J23107 promoter ready for insertion of a RBS/CDS insert
 +
|
 +
|
 +
|
 +
|}

Latest revision as of 18:11, 17 August 2007

yesterday -- tomorrow

Colony PCRs

The cloning finally seem to have worked, so we are doing colony PCRs to check if there is an insert and if it's got the right size.


Digestion reactions

The following digestion reactions (ON) were performed:


PURPLE = Vector
GREEN = Insert

Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 Size Description Date
D37 lox71-gfp(tripart) BI L12.2 miniprep products (from 30/07 ligation reaction) XbaI PstI lox71_RBS-gfp-double terminator extracted as a backward insert Aug. 16
Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 Size Description
D44 B0015 terminator BI B0015 in pSB1AK3 (MP3.3 Aug.14) XbaI Pst1 terminator B0015 extracted for use as a BI
D45 B0015 terminator FI B0015 in pSB1AK3 (MP3.4 Aug.14) EcoRI SpeI terminator B0015 extracted for use as a FI
D46 B0015 BV B0015 in pSB1AK3 (MP3.3 Aug.14) SpeI Pst1 B0015 in pSB1AK3 plasmid extracted for use as a BV
D47 pSB1A2 XP dig. pSB1A2 carrying Cre ORF (MP9.1) XbaI PstI pSB1A2 open vector ready for cloning of BI
D48 pSB1A2 ES dig. pSB1A2 carrying Cre ORF (MP9.2) EcoRI SpeI pSB1A2 open vector ready for cloning of FI
D49 lox71-gfptripart BI lox71-gfptripart in pSB1A2 (L12.2) XbaI Pst1 lox71-gfptripart extracted for use as a BI
D50 BBa_J61002 pTet SP dig. BBa_J61002 (MP13.1 Aug. 14) SpeI PstI BBa_J61002 pTet promoter vector ready for insertion of CDS as a BI
D51 RBS-dapAcoli BBinsert pTOPO of RBS-dapAcoli (MPT4.3) EcoRI Pst1 RBS-dapAcoli extracted for cloning as a biobrick
D52 RBS-dapAcoli BI pTOPO of RBS-dapAcoli (MPT4.3) XbaI Pst1 RBS-dapAcoli extracted for use as a BI
D53 RBS-dapAsubtilis BBinsert pTOPO of RBS-dapAsubtilis (MPT5.1) EcoRI Pst1 RBS-dapAsubtilis extracted for cloning as a biobrick
D54 RBS-dapAsubtilis BI pTOPO of RBS-dapAsubtilis (MPT5.1) XbaI Pst1 RBS-dapAsubtilis extracted for use as a BI
D55 RBS-dapAsubtilis FI pTOPO of RBS-dapAsubtilis (MPT5.2) EcoRI SpeI RBS-dapAsubtilis extracted for use as a FI



I was unable to perform the following digestion reactions (because their was no more MP1 or MP2 product left):


PURPLE = Vector
GREEN = Insert

Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 Size Description Date
D1 pJ23100 pJ23100 Spe1 Pst1 2096 Vector with the J23100 promoter ready for insertion of a RBS/CDS insert
D3 pJ23107 pJ23107 Spe1 Pst1 2096 Vector with the J23107 promoter ready for insertion of a RBS/CDS insert
Retrieved from "http://2007.igem.org/wiki/index.php/Paris/August_16"