Difference between revisions of "Part:BBa K274003:Experience"

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== '''Characterisation by Wits South Africa ''' ==
 
== '''Characterisation by Wits South Africa ''' ==
  
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We then decided to try and use the Dark Green E.chromi biobrick (BBa_K274003). Once again, although the part was excised using EcoR1 and Spe1 (Fermentas), when we attempted to clone anything in front of this part, by digestion with EcoR1 and Xba1 (Fermentas), we could not obtain any positive clones.  
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We attempted to try and use the Dark Green E.chromi biobrick (BBa_K274003) in our machine constructs. Although the part was excised using EcoR1 and Spe1 (Fermentas), when we attempted to clone anything in front of this part, by digestion with EcoR1 and Xba1 (Fermentas), we could not obtain any positive clones.  
  
 
Suspecting that perhaps the restriction sites were not present, we tested the plasmid samples through restriction digests.
 
Suspecting that perhaps the restriction sites were not present, we tested the plasmid samples through restriction digests.
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[[image: wits_xbal_digest.jpg|400px]]
 
[[image: wits_xbal_digest.jpg|400px]]
  
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'''Figure 2. E.chromi Dark Green digested with various restriction enzymes to confirm the presence of the Biobrick restriction sites.'''  
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'''Figure 1. E.chromi Dark Green digested with various restriction enzymes to confirm the presence of the Biobrick restriction sites.'''  
  
  
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As can be noted from Figure 2, the Xba1 site is not present in the Dark Green E.chromi Biobrick part, although the Spe1 and EcoR1 sites are present.
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As can be noted from Figure 1, the Xba1 site is not present in the Dark Green E.chromi Biobrick part, although the Spe1 and EcoR1 sites are present.
  
 
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Revision as of 21:39, 27 October 2010

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_K274003

User Reviews

E. coli turned dark green after transformation of this plasmid. This is unexpected as there is no promoter upstream of the operon.

BBa_K274003 Review No.2 UCL_London, Xiang Chen

Diagnostic DNA gel was run to test all 4 restriction sites. BBa_K274003 was cut with XbaI, PstI, EcoRI & SpeI, XbaI & PstI. A gel picture is shown as below.

This plasmid did not cut with Xba I restriction site in our hands. As such we could not use it to assemble after other parts.


K274003 Diagnostic Gel .jpg


Characterisation by Wits South Africa

We attempted to try and use the Dark Green E.chromi biobrick (BBa_K274003) in our machine constructs. Although the part was excised using EcoR1 and Spe1 (Fermentas), when we attempted to clone anything in front of this part, by digestion with EcoR1 and Xba1 (Fermentas), we could not obtain any positive clones.

Suspecting that perhaps the restriction sites were not present, we tested the plasmid samples through restriction digests.


Wits xbal digest.jpg

Figure 1. E.chromi Dark Green digested with various restriction enzymes to confirm the presence of the Biobrick restriction sites.


As can be noted from Figure 1, the Xba1 site is not present in the Dark Green E.chromi Biobrick part, although the Spe1 and EcoR1 sites are present.

UNIQddcd49946aff6031-partinfo-00000000-QINU UNIQddcd49946aff6031-partinfo-00000001-QINU