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Team:TU Delft/8 June 2010 content
From 2010.igem.org
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* Na2SO4 0.5g | * Na2SO4 0.5g | ||
* 1 ml of MT stock solution | * 1 ml of MT stock solution | ||
| + | * 1 ml of 1M MgSO4 | ||
* WATER 1L | * WATER 1L | ||
| Line 48: | Line 49: | ||
* MnCl2 0.19g | * MnCl2 0.19g | ||
* CoCl2 6H2O 2.8 g | * CoCl2 6H2O 2.8 g | ||
| - | * CaCl2 | + | * CaCl2 4H2O 1.84g |
* ZnSO4 7H2O 0.28 g | * ZnSO4 7H2O 0.28 g | ||
* CuSO4 0.16g | * CuSO4 0.16g | ||
| + | |||
| + | '''WARNING:''' Dear iGEM people be aware that the original E2 composition is the following (Lageveen ''et al.'', 1988): | ||
| + | |||
| + | * KH2PO4 3.7 g | ||
| + | * K2HPO4 7.5 g | ||
| + | * NaNH4HPO4.4H2O 3.5 g | ||
| + | * Citrate 5 g / 12 g | ||
| + | * 1 M MgSO4 . 7H2O 1 ml | ||
| + | * MT stock solution 1 ml | ||
| + | |||
| + | and the original MT stock solution composition is: | ||
| + | * FeSO4.7H2O 2.8 g | ||
| + | * MnSO4.H2O 2 g | ||
| + | * CoCl2.6H2O 2.8 g | ||
| + | * CaCl2.2H2O 1.48 g | ||
| + | * ZnSO4.7H2O 0.28 g | ||
| + | * CuCl2.2H2O 0.16 g | ||
Revision as of 10:25, 24 August 2010
Contents |
Eppy
Pieter just got a little present from Mariska van Ham :-) He's so happy with is eppy!
Today Nadine & Luke are learning how to work with HTML.
Lab work
BioBrick stocks
A number of Biobricks from the DNA distribution plates were suspended yesterday. Hugo and Kira did a transformation with 2 μL of the following BioBricks (antibiotic):
- pSB3T5 (Tetracycline)
- pSB1T3 (Tetracycline)
- pSB1C3 (Chloramphenicol)
- pSB3C5 (Chloramphenicol)
- pSB1K3 (Kanamycin)
- pSB1A3 (Ampicillin)
- I13401 (Ampicillin)
Because our own competent cells were not good enough we used commercially available competent Top10 cells (Invitrogen). The cells were plated on LB-agar with the appropriate antibiotic. Biobricks plated We will have to wait until tomorrow to see if they were successful or not. Hopefully we will see some colonies tomorrow. Wish us luck!
Thias PCRed some BioBricks with various primer concentrations and at different temperatures to see which PCR method would work best. The PCR products were tested on a gel, and we saw a difference between the primer concentrations and the temperatures used.
Media and Solutions
By Hugo
Nadine and I are planing to revive our Ps. putida strain (muahaha). So, we prepared some solid media for that purpose: LB agar and E2 agar. For LB medium this is the recipe that we use:
- Tryptone 10g
- Yeast extract 5 g
- NaCl 10 g
- Agar 15 g
- Water, as much as you need for 1 L of culture medium
For E2 medium this is the recipe that we use:
- (NH4)2HPO4 10 g
- K2HPO4 5 g
- Na2SO4 0.5g
- 1 ml of MT stock solution
- 1 ml of 1M MgSO4
- WATER 1L
This medium is really minimal, you can choose your own carbon source. Some literature recommend to use citrate, glucose, glycerol and, for our case, hydrocarbons. E2 medium is not complete without the following solution:
MT stock solution
- FeSO4 7H2O 2.8 g
- MnCl2 0.19g
- CoCl2 6H2O 2.8 g
- CaCl2 4H2O 1.84g
- ZnSO4 7H2O 0.28 g
- CuSO4 0.16g
WARNING: Dear iGEM people be aware that the original E2 composition is the following (Lageveen et al., 1988):
- KH2PO4 3.7 g
- K2HPO4 7.5 g
- NaNH4HPO4.4H2O 3.5 g
- Citrate 5 g / 12 g
- 1 M MgSO4 . 7H2O 1 ml
- MT stock solution 1 ml
and the original MT stock solution composition is:
- FeSO4.7H2O 2.8 g
- MnSO4.H2O 2 g
- CoCl2.6H2O 2.8 g
- CaCl2.2H2O 1.48 g
- ZnSO4.7H2O 0.28 g
- CuCl2.2H2O 0.16 g
