Registry/Measurement kit/Notebook/Design: Difference between revisions
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*4 - <bbpart>J04650</bbpart> | *4 - <bbpart>J04650</bbpart> | ||
*5 - <bbpart>B0032</bbpart> | *5 - <bbpart>B0032</bbpart> | ||
==Suggestions from peers== | |||
*Using LacZ with a Miller Assay to measure expression instead of an FP | |||
*3-series plasmid as the final vector (with flanking terminators) | |||
*Ensure appropriate spacing between RBS and coding region | |||
Revision as of 10:50, 15 June 2007
Design considerations for the expression measurement kit
Promoter
- Constituitive?
- Relative strength
- Location of transcription initiation (outside of or within the promoter?)
- Previous work
Ribosome Binding Site
- Relative Strength
- Previous work (<bbpart>B0032</bbpart>)
Fluorescent Protein
- Color?
- previous work (<bbpart>E0040</bbpart>)
- Tagged?
Terminator
- Do we need one?
- Which one?
- Previous work (<bbpart>B0015</bbpart>)
Preliminary Proposed Kit Designs
Promoter tester with GFP
- E0240 (pSB1A2)
Promoter tester with RFP
- B0032 (pSB1A2) + J04650 (pSB1AK3)
RBS tester with GFP
- R0040 (pSB1A2) + I13401 (pSB1A2)
RBS tester with RFP
- R0040 (pSB1A2) + J04650 (pSB1AK3)
Internal labels
- 1 - <bbpart>E0240</bbpart>
- 2 - <bbpart>R0040</bbpart>
- 3 - <bbpart>I13401</bbpart>
- 4 - <bbpart>J04650</bbpart>
- 5 - <bbpart>B0032</bbpart>
Suggestions from peers
- Using LacZ with a Miller Assay to measure expression instead of an FP
- 3-series plasmid as the final vector (with flanking terminators)
- Ensure appropriate spacing between RBS and coding region
