IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Implementation: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Ben Yi Tew (talk | contribs) m (→Maxi Prep) |
Ben Yi Tew (talk | contribs) m (→Maxi Prep) |
||
| Line 30: | Line 30: | ||
#Autoclave for 25 minutes | #Autoclave for 25 minutes | ||
#Wait until flask is hand-hot, you can hold the flask in your hand comfortably(~50°C) | #Wait until flask is hand-hot, you can hold the flask in your hand comfortably (~50°C) | ||
[Note: Please minimize contamination to the medium during preparation and after use. If LB us suspected to be contaminated, make a new one.] | [Note: Please minimize contamination to the medium during preparation and after use. If LB us suspected to be contaminated, make a new one.] | ||
Revision as of 03:04, 30 July 2007
Biofilm Detection: Implementation
Protocols
Maxi Prep
Summary: multiplying plasmid by putting them into the bacteria
Making LB Medium
To make 1 L of LB medium:
- 1 L distilled water
- 10 g tryptone
- 5 g yeast extract
- 5 g NaCl
- 1 ml 1 M NaOH
- Autoclave for 25 minutes
- Wait until flask is hand-hot, you can hold the flask in your hand comfortably (~50°C)
[Note: Please minimize contamination to the medium during preparation and after use. If LB us suspected to be contaminated, make a new one.]
==
- Add up to 100 µg/mL of ampicillin and/or 50 µg/mL of tetracyclin to LB
Electroporation
process of shocking the cells to make them take in the plasmid: plasmid naturally not diffusable
Plasmid Extraction
extraction of plasmid from the cell: breaking the cell and precipitate the plasmid
Restriction Digest
enzymes that cut on specific sites on the DNA
Ligation
sticking the DNA together
Agarose Electrophoresis
method to seperate DNA of different sizes, using an electric field, in a gel
DNA Extraction/Purification
extract the DNA from the gel
Biobricks
- Construct, clone and growth over 2 day period
