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Team:HokkaidoU Japan/Notebook/plastic Week 5
From 2012.igem.org
(Difference between revisions)
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==Transformation== | ==Transformation== | ||
<p> | <p> | ||
| - | Transformation of pGEM into BL21. This transformation is the ''' | + | Transformation of pGEM into BL21. This transformation is the '''bioplastic production test in BL21'''. |
#Added 1ul of plasmid DNA to 50ul of thawed competent cells on ice. | #Added 1ul of plasmid DNA to 50ul of thawed competent cells on ice. | ||
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#Plate 300ul of the transformation onto LBA dish and spread. | #Plate 300ul of the transformation onto LBA dish and spread. | ||
#Added 900ul of LB to 100ul of the transformation and plated 300ul of it onto LBA dish then spread. | #Added 900ul of LB to 100ul of the transformation and plated 300ul of it onto LBA dish then spread. | ||
| - | #Incubated the plates at 37C for | + | #Incubated the plates at 37C for 15hrs 30min(22:00~13:30). |
</p> | </p> | ||
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==August 3rd== | ==August 3rd== | ||
<div> | <div> | ||
| + | |||
==Liquid culture== | ==Liquid culture== | ||
<p> | <p> | ||
Liquid culture of Transformed BL21(DE3) | Liquid culture of Transformed BL21(DE3) | ||
| - | # | + | #Added 2ml of LBA into culture tubes. |
| - | # | + | #Resuspended 5 colonies. |
| - | # | + | #Incubated the tubes at 37C for OOhrs. |
</p> | </p> | ||
Revision as of 05:35, 3 August 2012
Contents |
August 2nd
Transformation
Transformation of pGEM into BL21. This transformation is the bioplastic production test in BL21.
- Added 1ul of plasmid DNA to 50ul of thawed competent cells on ice.
- Incubated on ice for 30min.
- Added 600ul of LB.
- Prepared and Labeled two petri dishes with LBA.
- Plate 300ul of the transformation onto LBA dish and spread.
- Added 900ul of LB to 100ul of the transformation and plated 300ul of it onto LBA dish then spread.
- Incubated the plates at 37C for 15hrs 30min(22:00~13:30).
August 3rd
Liquid culture
Liquid culture of Transformed BL21(DE3)
- Added 2ml of LBA into culture tubes.
- Resuspended 5 colonies.
- Incubated the tubes at 37C for OOhrs.
- Using agar stab***
We send out our part requests as agar stabs. The shelf life of these are short, so it is usually best to plate from the stab as soon as possible.
The agar should already have a hole from when it was stabbed. With an inoculating loop dipped into the stab, you can plate directly onto a petri dish with the appropriate antibiotic. Incubate the dish overnight at 37C (14-16hr) Pick a single colony to start up a culture Miniprep to extract plasmid DNA Use the part!
