IGEM:Peking/2007/Count-Conjugation-Procedure: Difference between revisions
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==Make more independent riboregulators== | ==Make more independent riboregulators== | ||
* We choose to use <bbpart> pSB1A3 < | * We choose to use <bbpart>pSB1A3</bbpart> and <bbpart>BBa_R0010</bbpart>(Plac) to express the taRNA, and <bbpart>pSB3K3</bbpart> and <bbpart>BBa_R0040</bbpart> to express <bbpart>BBa_E0040</bbpart>, whose translation is under the control of crRNA. | ||
* crRNA is synthesized as two partially overlap ~50bp long primer, and one cycle PCR can make it to a dsDNA with some BioBrick cloning sites flanked. | * crRNA is synthesized as two partially overlap ~50bp long primer, and one cycle PCR can make it to a dsDNA with some BioBrick cloning sites flanked. | ||
* crRNA should be cut by NotI at 5' and SpeI at 3' and cloned into pSB1A3 to get the full length BioBrick prefix and suffix. | * crRNA should be cut by NotI at 5' and SpeI at 3' and cloned into pSB1A3 to get the full length BioBrick prefix and suffix. | ||
* First, we will try out the UC Berkley riboregulators <bbpart> BBa_J23078 < | * First, we will try out the UC Berkley riboregulators <bbpart>BBa_J23078</bbpart> and <bbpart>BBa_J23066</bbpart> first. | ||
* This is the experimental [[IGEM:Peking/2007/Count-Procedure-Riboregulator | procedure]]. | * This is ''the exact experimental'' [[IGEM:Peking/2007/Count-Procedure-Riboregulator | ''procedure'']]. |
Revision as of 01:17, 9 July 2007
Make more independent riboregulators
- We choose to use <bbpart>pSB1A3</bbpart> and <bbpart>BBa_R0010</bbpart>(Plac) to express the taRNA, and <bbpart>pSB3K3</bbpart> and <bbpart>BBa_R0040</bbpart> to express <bbpart>BBa_E0040</bbpart>, whose translation is under the control of crRNA.
- crRNA is synthesized as two partially overlap ~50bp long primer, and one cycle PCR can make it to a dsDNA with some BioBrick cloning sites flanked.
- crRNA should be cut by NotI at 5' and SpeI at 3' and cloned into pSB1A3 to get the full length BioBrick prefix and suffix.
- First, we will try out the UC Berkley riboregulators <bbpart>BBa_J23078</bbpart> and <bbpart>BBa_J23066</bbpart> first.
- This is the exact experimental procedure.