Team:HKU-Hong Kong/Lab Diaries
From 2011.igem.org
(Difference between revisions)
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<LI>tetO2 – 0 – 3 was transformed to DH10b competent cells. | <LI>tetO2 – 0 – 3 was transformed to DH10b competent cells. | ||
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- | <LI>tetO2 – 0 –sfGFP | + | <LI>Production of tetO2 – 0 –sfGFP |
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+ | |style="font-family: georgia, helvetica, arial, sans-serif;font-size:2em;color:#01DF01;"|Lab Diaries | ||
+ | |- | ||
+ | |style="width:900px;"|'''Week 10''' | ||
+ | <OL> | ||
+ | <LI>EcoRI and PstI enzyme cut sites were added to both ends of HNS using PCR (for BioBricks construction). | ||
+ | <LI>Production of | ||
+ | <OL> | ||
+ | <LI>J23103RH | ||
+ | <LI>J23106RH; J23106RHH | ||
+ | <LI>J23116RH; J23116RHH | ||
+ | <LI>pET28a | ||
+ | <LI>pSB1C3-tdRema | ||
+ | </OL> | ||
+ | <LI>Checking fluorescence intensity of sfGFP and EGFP | ||
+ | <OL> | ||
+ | <LI>3 colonies from sfGFP streak plate and 1 colony from EGFP streak plate were picked. Each was inoculated into 0.5mL LB broth with 0.5uL Cm in 37C on rotary machine in warm room for around 3 hours. | ||
+ | <LI>Samples were then diluted 1:50 in fresh 2mL LB broth with 2uL Cm in 37C on rotary machine in warm room for around 3 hours. | ||
+ | <LI>Fluorescence intensity were checked at OD600 using spectrophotometer | ||
+ | </OL> |
Revision as of 17:48, 4 October 2011
Lab Diaries | |
Week 1
Transformation of reporter DNA (pEGFP-loxp-km-loxp) into DH10B (non-virulent strain E. coli) with antibiotic resistance (Chloramphenicol – Cm) | Lab Diaries |
Week 2
| Lab Diaries |
Week 3
| Lab Diaries |
Week 4-5
tetO2 – 0 and tetO2 – 4
| Lab Diaries |
Week 6
Overlap PCR was carried out to produce fusion protein gene
| Lab Diaries |
Week 7
Overlap PCR was carried out to produce fusion protein gene tetR::HNS (2-90)
| Lab Diaries |
Week 8
| Lab Diaries |
Week 9
| Lab Diaries |
Week 10
|