7/3/2023 0 Comments Noti hf neb![]() ![]() For Customers Outside of this Region Contact your local subsidiary or distributor. Links to this resource Related Products: NotI-HF To Request Technical Support Fill out our Technical Support Form. A subset of samples were gel-size-selected to remove adaptor-dimer bands on a 1% agarose gel (SeaKem) and purified using the column-based Gel Purification Kit (QIAGEN) and eluted in EB.įull paper Login or join for free to view the full paper. NotI-HF has been engineered with a mutation to exhibit significantly less star activity and comes supplied with rCutSmart Buffer. DNA was then purified using 45 μL SPRI beads. Ligase was heat-inactivated at 65☌ for 15 minutes followed by cooling on ice and addition of 1 μL 10× T4 DNA Ligase Buffer, 1 μL (1U) USER Enzyme (NEB), 1 μL (20U) NotI-HF (NEB), and 2 μL (10U) λ Exonuclease (NEB) and incubation at 37☌ for 2 hours. Two traditional DNA plasmids and two DNA minicircle constructs were evaluated. DNA was resuspended in 42 μL Elution Buffer (EB, QIAGEN) followed by addition of 5 μL 10× T4 DNA Ligase Buffer (NEB), 1 μL of each annealed adaptor (FT-½NotI and HP-½NotI), and 1 μL (400U) of T4 DNA Ligase (NEB) and incubated overnight at 16☌. We evaluated four DNA vaccine candidates for their ability to produce virus-like particles (VLPs) and elicit a protective immune response against Foot-and-mouth disease virus (FMDV) in cattle. 0.5 to 4 μg of Phi X 174 gDNA (Thermo Scientific) was restriction-digested using 5U of SspI (NEB) in 1× SspI Reaction Buffer for 2 hours at 37☌ to linearize DNA and produce blunt ends followed by SPRI bead purification. Thermo Scientific NotI restriction enzyme recognizes GCGGCCGC sites and cuts best at 37C in O buffer (Isoschizomers: CciNI). Phi X 174 nanopore libraries were constructed using a shotgun-ligation approach. ![]()
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