FokI restriction enzyme

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	3 jig RNA was converted to double-stranded cDNA with 200 U reverse transcriptase, and digested with 5 U of either of three class IIS restriction enzymes according to the supplier's protocol for 45 min. We used FokI (TaKaRa), BsmAI (NEB) and BsmFI (NEB). After phenol extraction and ethanol precipitation, the product was dissolved in 70 gl of distilled water.

Protocol tips
3 jig RNA was converted to double-stranded cDNA with 200 U reverse transcriptase, and digested with 5 U of either of three class IIS restriction enzymes according to the supplier's protocol for 45 min. We used FokI (TaKaRa), BsmAI (NEB) and BsmFI (NEB). After phenol extraction and ethanol precipitation, the product was dissolved in 70 gl of distilled water.

Publication protocol

"Total RNA was prepared from Balb/c mouse liver and kidneys as
described (2), and cDNA was produced as described by Gubler
Total RNA was prepared from Balb/c mouse liver and kidneys as described (2), and cDNA was produced as described by Gublerand Hoffman (3), using M-MLV reverse transcriptase (GibcoBRL), human placental RNase inhibitor (Toyobo), E.coli DNA polymerase I (Toyobo) and RNase H (Toyobo). The primer for the first strand cDNA synthesis was 5'-amino d(T)18. Usually3 jig RNA was converted to double-stranded cDNA with 200 U reverse transcriptase, and digested with 5 U of either of three class IIS restriction enzymes according to the supplier's protocol for 45 min. We used FokI (TaKaRa), BsmAI (NEB) and BsmFI (NEB). After phenol extraction and ethanol precipitation, the product was dissolved in 70 gl of distilled water."

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Manufacturer protocol

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