Generator

Part:BBa_K358006:Design

Designed by: Ken Kajita   Group: iGEM10_Kyoto   (2010-09-19)
Revision as of 09:09, 20 October 2010 by V08ken (Talk | contribs) (→‎References)

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].

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.