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Team:Newcastle/23 July 2010
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====Procedure==== | ====Procedure==== | ||
| - | ====Cell lysis==== | + | =====Cell lysis===== |
#Pellet cells by centrifugation at 3600rpm for 10 minutes | #Pellet cells by centrifugation at 3600rpm for 10 minutes | ||
#Pour off supernatant | #Pour off supernatant | ||
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#Heat sample for 30 mins mix every 5-10 mins | #Heat sample for 30 mins mix every 5-10 mins | ||
| - | ====RNAse treatment==== | + | =====RNAse treatment===== |
#Add 3 microlitres of RNAse A solution to the cell lysate | #Add 3 microlitres of RNAse A solution to the cell lysate | ||
#Mix by inverting 25 times and incubate at 37°C for 60 minutes | #Mix by inverting 25 times and incubate at 37°C for 60 minutes | ||
Revision as of 13:38, 23 July 2010
Contents |
Aims of the experiment
The aim of today's experiment is to extract genomic DNA from Bacillus subtilis strain 3610,genes from which will be needed for the swarming biobrick.
Materials Required
- Cells grown from yesterday
- Centrifuge
- pipette
- lysozyme
- Cell lysis solution
- RNAse solution
- protein orecipitation solution
- ice
- isopropanol
- ethanol
Procedure
Cell lysis
- Pellet cells by centrifugation at 3600rpm for 10 minutes
- Pour off supernatant
- Add 0.5ml of cell suspension solution, gently pipet up and down to resuspend and transfer to 1.5ml eppendorf tube
- Add 25microlitres of lysozyme and invert 25 times
- Incubate for 30minutes at 37°C inverting occasionally
- Centrifuge at 13000rpm for 10minutes to pellet the cells then remove the supernatant
- Add 0.5ml of cell lysis solution to the cell pellet and gently pipet up and down to lyse the cells
- Heat sample for 30 mins mix every 5-10 mins
RNAse treatment
- Add 3 microlitres of RNAse A solution to the cell lysate
- Mix by inverting 25 times and incubate at 37°C for 60 minutes
