System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay...
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System Engineering M2D3 11.03.09
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Transcript of System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay...
![Page 1: System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay for your mutants Part 3: sequence analysis.](https://reader035.fdocuments.net/reader035/viewer/2022081816/56649ebe5503460f94bc7974/html5/thumbnails/1.jpg)
System EngineeringM2D3
11.03.09
![Page 2: System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay for your mutants Part 3: sequence analysis.](https://reader035.fdocuments.net/reader035/viewer/2022081816/56649ebe5503460f94bc7974/html5/thumbnails/2.jpg)
Three part lab today Part 1: finish WesternPart 2: b-gal assay for your mutantsPart 3: sequence analysis
![Page 3: System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay for your mutants Part 3: sequence analysis.](https://reader035.fdocuments.net/reader035/viewer/2022081816/56649ebe5503460f94bc7974/html5/thumbnails/3.jpg)
Part 1: finish Western
Why are we performing a
Western blot?
![Page 4: System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay for your mutants Part 3: sequence analysis.](https://reader035.fdocuments.net/reader035/viewer/2022081816/56649ebe5503460f94bc7974/html5/thumbnails/4.jpg)
Part 2: b-gal assay
Why are we testing both light and dark
grown cells?
WT Cand 1 Cand 2 0
100200300400500600700800900
1000
lightdark
Miller units
Should we test + and – HA tag?
How many replicates should we test?
![Page 5: System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay for your mutants Part 3: sequence analysis.](https://reader035.fdocuments.net/reader035/viewer/2022081816/56649ebe5503460f94bc7974/html5/thumbnails/5.jpg)
Part 3: Sequence analysis
1.Retrieve the sequences
2. find the reverse complement
3.BLAST your sequence (= query) vs pCph8 (= subject)
Will pCph8 have HA tag?What will you do with your amino acid changes (if you have any)?
![Page 6: System Engineering M2D3 11.03.09. Three part lab today Part 1: finish Western Part 2: -gal assay for your mutants Part 3: sequence analysis.](https://reader035.fdocuments.net/reader035/viewer/2022081816/56649ebe5503460f94bc7974/html5/thumbnails/6.jpg)
Part 3: Sequence analysis