Team:HKU-Hong Kong/Lab Diaries

From 2011.igem.org

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|style="width:900px;"|'''Week 1'''
|style="width:900px;"|'''Week 1'''
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Transformation of reporter DNA (pEGFP-loxp-km-loxp) into DH10B (non-virulent strain E. coli) with antibiotic resistance (Chloramphenicol – Cm)
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|style="font-family: georgia, helvetica, arial, sans-serif;font-size:2em;color:#01DF01;"|Lab Diaries
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|style="width:900px;"|'''Week 2'''
<OL>
<OL>
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<LI>Transformation of reporter DNA (pEGFP-loxp-km-loxp) into DH10B (non-virulent strain E. coli) with antibiotic resistance (Chloramphenicol – Cm)</LI>
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<LI>tetO2 -1 (DNA binding site)
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<OL>
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<LI>Reverse PCR was used to insert tetO2 -1 into pEGFP-loxp-km-loxp (template DNA) by using forward and reverse primers which are with the tetO2 -1. By using reverse PCR, we can insert the tetO2 -1 site into the pEGFP while the plasmid produced is still in double-stranded circular form.
 +
<LI>pEGFP-loxp-km-loxp-tetO2-1 was then transformed into DH10B to greatly amplify the product by using bacterial cells
 +
</OL>
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<LI>tetO2 -2 (DNA biding site)
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<OL>
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<LI>Reverse PCR was used to insert tetO2 -2 into pEGFP-loxp-km-loxp
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<LI>2uL of 10-fold diluted pEGFP-loxp-km-loxp- tetO2 -2 was transformed into DH10B to greatly amplify the product by using bacterial cells
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</OL>
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<LI>tetO2 – 3 (DNA binding site)
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<OL>
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<LI>Reverse PCR was used to insert tetO2 – 3 into pEGFP-loxp-km-loxp
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<LI>2uL of 10-fold diluted pEGFP-loxp-km-loxp- tetO2 – 3 was transformed into DH10B to greatly amplify the product by using bacterial cells
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</OL>

Revision as of 16:33, 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
  1. tetO2 -1 (DNA binding site)
    1. Reverse PCR was used to insert tetO2 -1 into pEGFP-loxp-km-loxp (template DNA) by using forward and reverse primers which are with the tetO2 -1. By using reverse PCR, we can insert the tetO2 -1 site into the pEGFP while the plasmid produced is still in double-stranded circular form.
    2. pEGFP-loxp-km-loxp-tetO2-1 was then transformed into DH10B to greatly amplify the product by using bacterial cells
  2. tetO2 -2 (DNA biding site)
    1. Reverse PCR was used to insert tetO2 -2 into pEGFP-loxp-km-loxp
    2. 2uL of 10-fold diluted pEGFP-loxp-km-loxp- tetO2 -2 was transformed into DH10B to greatly amplify the product by using bacterial cells
  3. tetO2 – 3 (DNA binding site)
    1. Reverse PCR was used to insert tetO2 – 3 into pEGFP-loxp-km-loxp
    2. 2uL of 10-fold diluted pEGFP-loxp-km-loxp- tetO2 – 3 was transformed into DH10B to greatly amplify the product by using bacterial cells
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