Difference between revisions of "Part:BBa K358006:Design"

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===Design Notes===
 
===Design Notes===
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We performed PCR to get the lysis cassette[SRRz] and inserted before double terminator[BBa_B0015].
+
We had two steps of PCR to prepare this part.
 +
 
 +
 
 +
First, we performed PCR to get the lysis cassette[SRRz] and inserted before double terminator[BBa_B0015].
  
 
As the template, we used λ -''Hin''d Ⅲ digest[http://catalog.takara-bio.co.jp/PDFFiles/3403_DS_j.pdf].
 
As the template, we used λ -''Hin''d Ⅲ digest[http://catalog.takara-bio.co.jp/PDFFiles/3403_DS_j.pdf].
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Reverse primer:
 
Reverse primer:
 +
 +
 +
Then, we done the point mutation PCR.  This "λ -''Hin''d Ⅲ digest" isn't the wt-DNA and there is a mutation on S gene.
 +
 +
By doing this PCR, we prepared wt-SRRz gene.
  
 
===Source===
 
===Source===

Revision as of 08:55, 20 October 2010

lambda lysis cassette with terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We had two steps of PCR to prepare this part.


First, we performed PCR to get the lysis cassette[SRRz] and inserted before double terminator[BBa_B0015].

As the template, we used λ -Hind Ⅲ digest[http://catalog.takara-bio.co.jp/PDFFiles/3403_DS_j.pdf].

Forward primer:


Reverse primer:


Then, we done the point mutation PCR. This "λ -Hind Ⅲ digest" isn't the wt-DNA and there is a mutation on S gene.

By doing this PCR, we prepared wt-SRRz gene.

Source

Used λ -Hind digest and BBa_B0015.

References

C. Chang, K. Nam, and R. Young, “S gene expression and the timing of lysis by bacteriophage lambda,” J. Bacteriol., vol. 177, Jun. 1995, pp. 3283-3294.[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC177022/pdf/1773283.pdf]