QIAEX II Gel Extraction Kit (150)

Protocol tips

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	Follow manufacturer's instructions

Protocol tips
Follow manufacturer's instructions

Publication protocol

Step 1. Assembly of Hexamers. 1. Pick six TAL RVD plasmids, 200-250 ng for each, 200 ng pTemp-S.2. Add 2 µl 10x T4 DNA ligase buffer, 1 ul BsmBI (New England Biolabs, R0580), andmake up to 20 µl with ddH2O. 3. Incubation 60 minutes at 55°C.4. Add 1 µl T4 DNA ligase (New England Biolabs, M0202). Then run cycle as: 37° C 5min,16 °C 10 min, total 9 cycles, followed by 16 °C 20 min . 5. Add Plasmid-Safe nuclease (Epicentre, E3101K) 1 µl into each reaction, then incubate30-60 min at 37° C. Take out 6 µl of each ligation product and save at -20 °C. The rest ofthree hexamer ligation products for each side TALEN are combined into ONE tube.6. Add 8 µl QIAEX II Suspension into each tube and purify through QIAEX II Gel ExtractionKit (Qiagen) following the manual.7. Elute DNAs in 13 µl EB buffer (~11.5 µl may be recovered)

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Manufacturer protocol

Download the product protocol from Qiagen for QIAEX II Gel Extraction Kit (150) below.

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