From 2010.igem.org
(Difference between revisions)
|
|
| Line 22: |
Line 22: |
| | ==July, 6th== | | ==July, 6th== |
| | | | |
| | + | We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection. Inoculum from glycerol stpcks (8ul in 5ml LB) for: |
| | + | *<partinfo>BBa_J23101</partinfo> |
| | + | *<partinfo>BBa_J23110</partinfo> |
| | + | *<partinfo>BBa_J23116</partinfo> |
| | + | *<partinfo>BBa_J23118</partinfo> |
| | + | *<partinfo>BBa_J23114</partinfo> |
| | + | *<partinfo>BBa_B0033</partinfo> |
| | + | |
| | + | TECAN experiment started at 10:20am. |
| | + | |
| | + | In the morning one single colony was peaked for BW53474 E. coli strain and glycerol stock was prepared after about 7 hours (37°C 220rpm) |
| | + | |
| | + | Inoculum of: |
| | + | *BW53474 (falcon containing culture for glycerol stock was re-filled with 5ml LB) |
| | + | *MG1655 (from glycerol stock) |
| | + | *BW25141 (from glycerol stock) |
| | + | *BW25142 (from glycerol stock) |
| | | | |
| | ==July, 7th== | | ==July, 7th== |
Revision as of 13:14, 26 July 2010
|
|
|
|
|
JULY: WEEK 2
July, 5th
LB agar plates prepared:
- LB+Amp 100 (500ml)
- LB+Cm 34 (500ml)
- LB (250ml)
- LB+Cm 12.5 (250ml)
July, 6th
We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection. Inoculum from glycerol stpcks (8ul in 5ml LB) for:
- <partinfo>BBa_J23101</partinfo>
- <partinfo>BBa_J23110</partinfo>
- <partinfo>BBa_J23116</partinfo>
- <partinfo>BBa_J23118</partinfo>
- <partinfo>BBa_J23114</partinfo>
- <partinfo>BBa_B0033</partinfo>
TECAN experiment started at 10:20am.
In the morning one single colony was peaked for BW53474 E. coli strain and glycerol stock was prepared after about 7 hours (37°C 220rpm)
Inoculum of:
- BW53474 (falcon containing culture for glycerol stock was re-filled with 5ml LB)
- MG1655 (from glycerol stock)
- BW25141 (from glycerol stock)
- BW25142 (from glycerol stock)
July, 7th
July, 8th
Transformation of 1ul (~4ng) of miniprepped <partinfo>BBa_J23118</partinfo> into 100ul of home-made competent cells
- MG1655
- BW25141
- BW25142
- BW53474
- DH5-alpha (as control)
to verify the transformation efficiency.
Transformed bacteria were plated on LB+Amp and left overnight in oven, 37°C.
July, 9th
We checked the presence of colonies in plates.
All plates showed red colonies!
 MG1655 plate (with satellite colonies) | |
|
| |
|
We calculated (thanks Nicolò) efficiency as #colonies/ug DNA plated
| Strain | #colonies | Efficiency
|
| MG1655 | 379 | ~10^5
|
| BW25141 | 1431 | ~10^6
|
| BW25142 | 2374 | ~10^6
|
| BW53474 | 9984 | ~10^7
(very difficult to count correctly)
|
| DH5alpha | 7475
(only 100ul of cells plated) | ~ 10^7
(very difficult to count correctly,
previous tests showed an efficiency of ~10^8)
|
|
|
|
"
