Difference between revisions of "Part:BBa K1498001:Experience"
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===Applications of BBa_K1498001===
 
===Applications of BBa_K1498001===
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<img class="leftAlignImg"src="https://static.igem.org/mediawiki/2014/e/e7/Rhodamineb_Sheffield2014.png" style="float:left;height:300px">
  
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<img class="leftAlignImg"src="https://static.igem.org/mediawiki/2014/1/19/Transilluminator_Sheffield2014.png" style="float:left;height:300px;padding:0px 10px 0px 0px"> <!-- Lipase plate, UV-->
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Lipase activity of secreted enzyme was shown on a 1.5% agar plate, containing 0.02% w/v rhodamine B and 6.6% v/v olive oil. The sterile filtered supernatant of DH5a E. coli cells expressing BBa_K258006 under the control of BBa_J23110 and the ABC transporter (BBa_K258008) was applied to wells in the rhodamine B plates. Sterile Luria-Bertani broth and the supernatant of cells expressing BBa_J23110 only were loaded as negative controls. Following incubation at 37°c overnight a halo was seen around the lipase containing supernatant. Under UV light fluorescent halo was observed.  
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'''Rhodamine B assay'''
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Lipase activity of secreted enzyme was shown on a 1.5% agar plate, containing 0.02% w/v rhodamine B and 6.6% v/v olive oil. The sterile filtered supernatant of cells expressing BBa_K258006 under the control of BBa_J23110 and the ABC transporter (BBa_K258008) was applied to wells in the rhodamine B plates. Sterile Luria-Bertani broth and the supernatant of cells expressing BBa_J23110 only were loaded as negative controls. Following incubation at 37°c overnight a halo was seen around the lipase containing supernatant (See left hand image: left: secreted lipase, middle:BBa_J23110 only, right: LB broth only . Under UV light fluorescent halo was observed (see right hand image: left: LB broth only, middle:BBa_J23110 only, right: secreted lipase.)
  
 
'''Reference.'''
 
'''Reference.'''

Latest revision as of 03:10, 18 October 2014

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Applications of BBa_K1498001

Rhodamine B assay

Lipase activity of secreted enzyme was shown on a 1.5% agar plate, containing 0.02% w/v rhodamine B and 6.6% v/v olive oil. The sterile filtered supernatant of cells expressing BBa_K258006 under the control of BBa_J23110 and the ABC transporter (BBa_K258008) was applied to wells in the rhodamine B plates. Sterile Luria-Bertani broth and the supernatant of cells expressing BBa_J23110 only were loaded as negative controls. Following incubation at 37°c overnight a halo was seen around the lipase containing supernatant (See left hand image: left: secreted lipase, middle:BBa_J23110 only, right: LB broth only . Under UV light fluorescent halo was observed (see right hand image: left: LB broth only, middle:BBa_J23110 only, right: secreted lipase.)

Reference.

Kouker, G., & Jaeger, K. E. (1987). Specific and sensitive plate assay for bacterial lipases. Applied and environmental microbiology, 53(1), 211–3

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