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

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===Design Notes===
 
===Design Notes===
We performed PCR to get the lysis cassette[SRRz] and inserted before double terminator[BBa_B0015].
+
[[image:K358006.jpg]]
 +
 
 +
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].
  
Forward primer:  
+
Fwd primer:
 +
 
 +
Rev 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.
 +
 
 +
Fwd primer:
 +
 
 +
Rev primer:
  
  
Reverse primer:
+
See details in notebook.
  
 
===Source===
 
===Source===
  
BBa_K358004 and BBa_B0015
+
Used λ -''Hin''d digest and BBa_B0015.
  
 
===References===
 
===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.

Latest revision as of 11:42, 23 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

K358006.jpg

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].

Fwd primer:

Rev 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.

Fwd primer:

Rev primer:


See details in notebook.

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.