Difference between revisions of "Part:BBa K1323008:Experience"
 
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[[File:sRNA Graph.png]]
 
[[File:sRNA Graph.png]]
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== Legend ==
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YFP cassette (pSB3K3 backbone, sarA protomer [https://parts.igem.org/Part:BBa_K1323021 BBa_K1323021],TIR RBS [https://parts.igem.org/Part:BBa_K1323016 BBa_K1323016], YFP cds [https://parts.igem.org/Part:BBa_K1323010 BBa_K1323010])
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sRNA 2 ([https://parts.igem.org/Part:BBa_K1323006 BBa_K1323006])
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sRNA 3 ([https://parts.igem.org/Part:BBa_K1323007 BBa_K1323007])
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Control sRNA
  
  
 
==sRNAi Silencing assay ==
 
==sRNAi Silencing assay ==
  
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YFP construct (pSB3K3 backbone, sarA protomer [https://parts.igem.org/Part:BBa_K1323021 BBa_K1323021],TIR rbs [https://parts.igem.org/Part:BBa_K1323016 BBa_K1323016], YFP cds[https://parts.igem.org/Part:BBa_K1323010 BBa_K1323010] and sRNA constructs were co-transformed into DH5α. Colonies were streak purified and grown in LB supplemented with ampicillin. A restriction digest was run to confirm that both plasmids were in the colonies selected. Overnight cultures were prepared in triplicate for each sample. 50 µL of overnight culture was used to inoculate fresh media, and once the OD600 reached 0.5-0.6, the YFP fluorescence was measured.  
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The YFP cassette and sRNA constructs were co-transformed into DH5α. The control for YFP expression included a sample with only the YFP cassette (no sRNA constructs), and a control sRNA that does not specifically target YFP. Colonies were streak purified and grown in LB supplemented with ampicillin. A restriction digest was run to confirm that both plasmids were in the colonies selected for the antibiotic resistance tests. Overnight cultures, in biological triplicate for each sample, were prepared. 50 µL of overnight culture was used to inoculate fresh media, and once the OD600 reached 0.5-0.6, the YFP fluorescence was measured.
 
===Applications of BBa_K1323008===
 
===Applications of BBa_K1323008===
  

Latest revision as of 20:46, 17 October 2014


SRNA Graph.png


Legend

YFP cassette (pSB3K3 backbone, sarA protomer BBa_K1323021,TIR RBS BBa_K1323016, YFP cds BBa_K1323010)

sRNA 2 (BBa_K1323006)

sRNA 3 (BBa_K1323007)

Control sRNA


sRNAi Silencing assay

The YFP cassette and sRNA constructs were co-transformed into DH5α. The control for YFP expression included a sample with only the YFP cassette (no sRNA constructs), and a control sRNA that does not specifically target YFP. Colonies were streak purified and grown in LB supplemented with ampicillin. A restriction digest was run to confirm that both plasmids were in the colonies selected for the antibiotic resistance tests. Overnight cultures, in biological triplicate for each sample, were prepared. 50 µL of overnight culture was used to inoculate fresh media, and once the OD600 reached 0.5-0.6, the YFP fluorescence was measured.

Applications of BBa_K1323008

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