Paris/August 16
From IGEM07
< Paris
[edit]
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.
[edit]
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 | |||
