Paris/August 16
From 2007.igem.org
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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 | |||
