NcoI restriction enzyme

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Plasmid pGFPuv4-NF, which contains a non-fluorescent GFP gene coding sequence, was prepared by restriction digestion with NcoI (Takara) followed by treatment of pGFPuv4 (Ito et al., 1999) with T4 DNA polymerase (Takara) and re-ligation with T4 DNA ligase (Takara).

Protocol tips
Plasmid pGFPuv4-NF, which contains a non-fluorescent GFP gene coding sequence, was prepared by restriction digestion with NcoI (Takara) followed by treatment of pGFPuv4 (Ito et al., 1999) with T4 DNA polymerase (Takara) and re-ligation with T4 DNA ligase (Takara).

Publication protocol

Plasmid pGFPuv4-NF, which contains a non-fluorescent GFP gene coding sequence, was prepared by restriction digestion with NcoI (Takara) followed by treatment of pGFPuv4 (Ito et al., 1999) with T4 DNA polymerase (Takara) and re-ligation with T4 DNA ligase (Takara). The expression vector was prepared from plasmid pGFPuv4-NF by PCR using GFP-D1 and GFP-D2 (10 pmol each, Table II) as primers. These primers contain AatII and NspV restriction sites and are designed so as not to modify the open reading frame of the GFP gene. PCR assays consisted of 30 cycles of denaturation (94°C, 30 s), annealing (55°C, 30 s) and extension (72°C, 2.5 min) after pre-denaturation (94°C, 2 min). The PCR products were finally treated with AatII (New England Biolabs) and NspV (Takara) and purified using a QIAquick column (QIAGEN, Hilden, Germany)

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Restriction Enzymes NcoI using NcoI restriction enzyme from Takara Bio Inc.

Manufacturer protocol

Download the product protocol from Takara Bio Inc for NcoI restriction enzyme below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Restriction Enzymes NcoI using NcoI restriction enzyme from Takara Bio Inc. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.