Difference between revisions of "Part:BBa B0015:Experience"

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The transformation, Plasmid miniprep and gel (undigested and digested with Xbal and PstI) worked as expected.
 
The transformation, Plasmid miniprep and gel (undigested and digested with Xbal and PstI) worked as expected.
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Revision as of 02:10, 4 October 2012

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_B0015

  • PCR Problems: Using primers VR and VF2 to PCR B0010 results in excess bands. VR can anneal to B0010, resulting in shorter bands than expected. A full description of the problem is available here.
  • We used this piece to terminate many of our Berkeley iGEM 2005 Parts. It appeared to work quite well. [Smelissali 6/7/06]
  • There have been reports from the Davidson iGEM team that the plasmid from the iGEM 2006 Repository for this part did NOT contain this biobrick. If you are experiencing this or a similar concern, please contact either myself or [http://2006.igem.org/User:Meaganl Meagan Lizarazo] [--Smelissali 14:26, 2 August 2006 (EDT)]

Issues in using this as vector

  • The standard assembly described here gives very poor results when whatever part BBa_Xxxxx is to be put in front of BBa_B0015 (< 5 cfu/plate). The procedure described below gave much better results (> 200 cfu/plate):
    • Cut BBa_B0015 with XbaI and PstI.
    • Cut BBa_Xxxxx with SpeI and PstI.
    • Ligate the two parts.
Bmoeyaert 08:50, 4 September 2008 (UTC)

User Reviews

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Antiquity

This review comes from the old result system and indicates that this part worked in some test.

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Aberdeen_Scotland 2009

The transformation, Plasmid miniprep and gel (undigested and digested with Xbal and PstI) worked as expected.

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Michigan iGEM 2012

As previously observed by other teams, when BBa_B0015 is PCR amplified using VF2 and VR unexpected bands of varying lengths (a bright band at 500bp and several dim ones below 500bp) appear on the electrophoresis gel.

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