IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/08/15

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(Floor Two)
Current revision (09:46, 22 August 2013) (view source)
(Floor Two)
 
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==Floor One==
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Made 4 x 500 ml and 2 x 1000 ml of SFM + Ny.
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Completed more lawns of single Streptomyces colonies.
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Decided to no longer re-streak contaminated plates as we have sufficient plates growing to produce a reasonable no. of spore stocks.
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==Floor Two==
==Floor Two==
The two plates and control plate from the previous day were successful, so two colonies were picked from plate one and used to inoculate two 10ml LB solutions and left to incubate overnight at 37°C. <br>
The two plates and control plate from the previous day were successful, so two colonies were picked from plate one and used to inoculate two 10ml LB solutions and left to incubate overnight at 37°C. <br>

Current revision

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Floor One

Made 4 x 500 ml and 2 x 1000 ml of SFM + Ny.

Completed more lawns of single Streptomyces colonies.

Decided to no longer re-streak contaminated plates as we have sufficient plates growing to produce a reasonable no. of spore stocks.


Floor Two

The two plates and control plate from the previous day were successful, so two colonies were picked from plate one and used to inoculate two 10ml LB solutions and left to incubate overnight at 37°C.
The sequencing data of the analysed biobricks were studied further. Sequence 1,2,5, and 6 all matched their expected DNA Sequences. Sequence 3 didn't match the expected sequence and sequence 4 contained extra NdeI restriction sites.
Two LB solutions were inoculated with BL21 PlySs and BL21 Rosetta PlySs seperately and incubated overnight at 37°C.

Fig.1:Transformation of pETAntA plasmid into BL21 E.Coli cells and spread onto LB agar + Kanamycin plates.
Fig.1:Transformation of pETAntA plasmid into BL21 E.Coli cells and spread onto LB agar + Kanamycin plates.



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