Difference between revisions of "Template:Loop/Universal Acceptor"

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Latest revision as of 16:50, 9 July 2019

Universal Acceptor

When BBa_J04452 and pSB1C00 are paired together, pSB1C00 can be used as a Universal Acceptor to clone in new Level 0 basic parts made through PCR or synthesis. View the BBa_J04452's Get This Part page to find a sample in the distribution kit.


PSB1C00-J04452.png


1. Add the following ends to your part sample through PCR or synthesis. This will result in your part flanked by the desired Type IIS fusion sites (numbered) and Sap1 sites.

PCR_for_pSB1C00.png


2. When both your part sample and the whole acceptor plasmid (pSB1C00-BBa_J04452) are cut with Sap1, the RFP device will be excised, leaving a partial Bsa1 sequence. Your cut part sample will have the correct sticky ends to ligate in.

PCR_and_pSB1C00_cut.png


3. Ligated product will result in your part in the backbone and flanked by Bsa1 sites. You can select for your liagted part by screening against the RFP device.

Part_in_pSB1C00.png


More details to follow.