Difference between revisions of "Part:BBa K1321334:Experience"
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===Applications of BBa_K1321334===
 
===Applications of BBa_K1321334===
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This part was cloned into part  BBa_K1321333 by Biobrick cloning. The destination vector, containing the AraC-pBAD regulatory elements, was linearized using SpeI and PstI and AcsAB (previously digested with XbaI and PstI, was ligated at a 1:1 using the T4 ligase. The ligation mix was transformed into chemically competent DH10B Escherichia coli and plated on LB Agar+Chloramphenicol plates, grown overnight at 37 degrees celsius. 5 ml LB falcons supplied with 50 ug/ml Chloramphenicol were inoculated with a selection of freshly grown colonies for further restriction analysis (refer to Results for additional information)
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This part was cloned into part  BBa_K1321333 by Biobrick cloning, to yield BBa_K1321336. The destination vector, containing the AraC-pBAD regulatory elements, was linearized using SpeI and PstI and AcsAB (previously digested with XbaI and PstI, was ligated at a 1:1 using the T4 ligase. The ligation mix was transformed into chemically competent DH10B Escherichia coli and plated on LB Agar+Chloramphenicol plates, then incubated overnight at 37 degrees. 5 ml LB falcons supplied with 50 ug/ml Chloramphenicol were inoculated with a selection of freshly grown colonies for further restriction analysis (refer to Results for additional information).
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The functionality of part BBa_K1321336 was assayed by inducing AcsAB-containing cells with 0.1% Arabinose in 5mL LB supplied with 1% Glucose and Chloramphenicol. Overnight incubations were set up at 30 °C and 37 °C, and both empty vector controls and un-induced controls were also assayed. During sonication procedures LB, soluble and non-soluble fractions (the latter dissolved in 1x PBS) were kept for further analysis. Congo Red at a concentration of 20uM was added to all samples, which were further incubated for 2h at static conditions and at room temperature to allow for Congo Red binding.
  
 
===User Reviews===
 
===User Reviews===

Revision as of 10:23, 21 October 2014

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1321334

This part was cloned into part BBa_K1321333 by Biobrick cloning, to yield BBa_K1321336. The destination vector, containing the AraC-pBAD regulatory elements, was linearized using SpeI and PstI and AcsAB (previously digested with XbaI and PstI, was ligated at a 1:1 using the T4 ligase. The ligation mix was transformed into chemically competent DH10B Escherichia coli and plated on LB Agar+Chloramphenicol plates, then incubated overnight at 37 degrees. 5 ml LB falcons supplied with 50 ug/ml Chloramphenicol were inoculated with a selection of freshly grown colonies for further restriction analysis (refer to Results for additional information).

The functionality of part BBa_K1321336 was assayed by inducing AcsAB-containing cells with 0.1% Arabinose in 5mL LB supplied with 1% Glucose and Chloramphenicol. Overnight incubations were set up at 30 °C and 37 °C, and both empty vector controls and un-induced controls were also assayed. During sonication procedures LB, soluble and non-soluble fractions (the latter dissolved in 1x PBS) were kept for further analysis. Congo Red at a concentration of 20uM was added to all samples, which were further incubated for 2h at static conditions and at room temperature to allow for Congo Red binding.

User Reviews

UNIQ26d8f6a2367b44d9-partinfo-00000000-QINU UNIQ26d8f6a2367b44d9-partinfo-00000001-QINU