Editing GoldenGate TALEN Assembly (section)

=== <br>DAY5<br><br>  ===

19. Miniprep the pTAL vectors containing your final full-length TALEN<br><br> 

20. Optional restriction digestion testing/sequencing: 

<br>Use enzymes BstAPI (or '''StuI''') and '''AatII''' to cut out the final array of repeats –check on a gel.&nbsp; You can use BspEI enzyme, which cuts only in HD repeats (except HD1 repeats) – on the gel you’ll get a pattern resembling your TALEN sequence and HD.<br> 

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21. Your TALEN is ready to use in a yeast-based DNA cleavage assay. Or further cloning, you can cut the TALE domain out using BamHI or combination of XbaI and EcoRV (blunt) repeats position 

<br>Note: there is no BspEI site in the first (i.e. #1) and the last (i.e. #N) HD repeat and sequence with primers SeqTALEN 5-1 and TAL_R2<br><br> 

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