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]. | ||
| − | + | 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: | ||
| − | + | See details in notebook. | |
===Source=== | ===Source=== | ||
| − | + | 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.