Difference between revisions of "Part:BBa P1010:Experience"

Revision as of 06:28, 18 October 2009

Applications of BBa_P1010

This is the part that is present inside the BioBricks insertion site in many of the pSB series of plasmids. It is sufficient for positive selection even in pSB2* series plasmids in the absence of inducer (~4-5 copies per cell). During assembly work at the Registry, we have found that point mutations (particularly frame shifts) occur at a high enough rate that several colonies will survive during a normal transformation. -Randy

User Reviews

UNIQ95b9f37bc9d9b917-partinfo-00000000-QINU

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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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iGEM Groningen 2009

P1010 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or self ligated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result. The ccdB cell death gene worked as expected killing our E. coli TOP10 cells, and keeping our E. coli DB3 cells alive. After noticing the inconsistent sequencing result for the pSB2K3 plasmid with ccdB cell death gene, we decided to choose a different pSB2K3 plasmid with random part to continue with our assemblies. This to minimize the chance of unwanted surprises in the end.

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 This review comes from the old result system and indicates that this part worked in some test.
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+{|width='80%' style='border:1px solid gray'
+|-
+|width='10%'|
+<partinfo>BBa_P1010 AddReview 4</partinfo>
+<I>iGEM Groningen 2009</I>
+|width='60%' valign='top'|
+P1010 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or self ligated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result. The ccdB cell death gene worked as expected killing our ''E. coli'' TOP10 cells, and keeping our ''E. coli'' DB3 cells alive. After noticing the inconsistent sequencing result for the pSB2K3 plasmid with ccdB cell death gene, we decided to choose a different pSB2K3 plasmid with random part to continue with our assemblies. This to minimize the chance of unwanted surprises in the end.
 |}
 <!-- DON'T DELETE --><partinfo>BBa_P1010 EndReviews</partinfo>