Fastdigest ecorv

Manufactured by Thermo Fisher Scientific

The FastDigest EcoRV is a restriction enzyme produced by Thermo Fisher Scientific. It recognizes and cleaves the DNA sequence 5'-GAT▼ATC-3' and its reverse complement 5'-CTA▼TAG-3'. The enzyme is designed for fast and efficient DNA digestion, providing complete digestion in as little as 5-15 minutes at 37°C.

Automatically generated - may contain errors

Lab products found in correlation

Nebnext ultra 2 end repair da tailing module, by New England Biolabs (1 mentions) Rnase 1, by Thermo Fisher Scientific (1 mentions) Nebnext ffpe dna repair mix, by New England Biolabs (1 mentions) Fastdigest mssi, by Thermo Fisher Scientific (1 mentions) Blunt ta ligase master mix, by New England Biolabs (1 mentions) Fastdigest drai, by Thermo Fisher Scientific (1 mentions) Rabbit anti vinculin, by Cell Signaling Technology (1 mentions) Fastdigest bamhi, by Thermo Fisher Scientific (1 mentions) Cag cas9 t2a egfp ires puro, by Addgene (1 mentions) Nebnext quick ligation reaction buffer, by New England Biolabs (1 mentions) High prime dna labeling kit, by Roche (1 mentions) Phusion high fidelity dna polymerase, by Thermo Fisher Scientific (1 mentions) Pbluescript 2 ks, by Addgene (1 mentions) T4 dna ligase, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

FastDigest (88 citations) EcoRV (26 citations)

2 protocols using fastdigest ecorv

CRISPR-Cas9 Genome Editing Workflow

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies: anti-MGME1 antibody (Sigma, HPA040913), rabbit-anti-vinculin (Cell Signaling, #13901), goat-anti-rabbit lgG HRP (Antibodies online, ABIN101744).
Recombinant constructs: CAG-Cas9-T2A-EGFP-ires-puro (Addgene # 78311).
Enzymes: FastDigest EcoRV (Thermo Scientific FD0303), FastDigest DraI (Thermo Scientific FD0224), FastDigest BamHI (Thermo Scientific FD0054), FastDigest MssI (Thermo Scientific FD1344), FastDigest Alw44I (Thermo Scientific FD0044), TopoIV (Inspiralis T4001), RNaseI (Thermo Scientific EN0601), T4 ligase and NEBNext Quick Ligation Reaction Buffer (New England Biolabs B6058).
Kits: High Prime DNA Labeling Kit (Roche 11585584001), NEBNext FFPE DNA Repair Mix (New England Biolabs #M6630), NEBNext Ultra II End repair/dA-tailing Module (New England Biolabs #E7546), Blunt/TA Ligase Master Mix (New England Biolabs M0367).

Fragkoulis G., Hangas A., Fekete Z., Michell C., Moraes C.T., Willcox S., Griffith J.D., Goffart S, & Pohjoismäki J.L. (2024). Linear DNA-driven recombination in mammalian mitochondria. Nucleic Acids Research, 52(6), 3088-3105.

+ Open protocol

+ Expand

Plasmid Construction Using GoldenGate and Hi-Fi Cloning

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Plasmids were built using both GoldenGate and Hi-Fi cloning [113 (link), 114 (link)]. Sequences and annotations are provided in GenBank format on the Additional file 3. Briefly, DNA fragments were PCR-amplified using Phusion High-Fidelity DNA polymerase (Thermo Scientific). Primers used in PCR reactions for GoldenGate were designed to add BsaI sites in both extremities of the amplicon while the primers for HiFi cloning were designed to include a complementary overhang in their extremities [114 (link)]. Primer sequences and construction schemes for each plasmid are available from the corresponding authors, RPO and JTM, upon reasonable request. All PCR products were cloned into a modified pBluescriptII KS( +) (Addgene #62540) lacking BsaI restriction site in a one-step digestion/ligation using FastDigest SmaI or FastDigest EcoRV (Thermo Scientific) and T4 DNA-ligase (Invitrogen) with 10 mM ATP. Final plasmid assembly by GoldenGate cloning was conducted as described previously [114 (link)]. Successful cloning was confirmed by Sanger sequencing (GATC Biotech).

Estevez-Castro C.F., Rodrigues M.F., Babarit A., Ferreira F.V., de Andrade E.G., Marois E., Cogni R., Aguiar E.R., Marques J.T, & Olmo R.P. (2024). Neofunctionalization driven by positive selection led to the retention of the loqs2 gene encoding an Aedes specific dsRNA binding protein. BMC Biology, 22, 14.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!