BsaI‐HF‐v2 restriction enzyme and 10× CutSmart buffer (NEB R3733) DNA spin‐purification kit (e.g., NEB T1030 or Zymo Research D4003) Custom oligonucleotides (Top and Bottom, each 100 µM; see Basic Protocol 1, Strategic Planning, and Fig. BsaI-HFv2. 野生型 BsaI および BsaI-HF の改良型酵素である。認識配列などの特異性はそのままで、バッファー適合性が大きく向上している。また、反応性の向上により、GoldenGate法に適している。 由来： Bacillus stearothermophilus 6-55 (Z. Chen)からクローニングされたBsaI遺伝子の遺伝子改変型(Y231F)を有する … HF® (High-Fidelity) R3733L R3733S. Golden Gate Assembly Workflow Find out how Golden Gate Assembly can be used to quickly join multiple DNA fragments. This reduces the reaction time to ca. Nucleic Acid Reagents Restriction Enzymes. Locate commercially available restriction enzymes by category, name, recognition sequence, or overhang. Ineligible item added to cart. Is there a difference in cutting close to the ends between BsaI-HF and BsaI? BsaI-HFv2. 76284-692EA 294.18 USD. Therefore, Type IIS REases that create 4-base overhangs (such as BsaI/BsaI-HF®v2, BbsI, BsmBI, Esp3I and PaqCI) are preferred. *included in NEB ® Golden Gate Assembly Kit (BsaI-HF ® v2) Reaction set-up. First, a volume of 0.5 µl of the respective restriction enzyme BbsI (5 units; ThermoFisherScientific, Waltham, US) or BsaI-HF®v2 (10 units; … 5000 units-1 + Unavailable in your region ® ® ® ® ® ® ®. Your personal data will be used to support your experience throughout this website, to manage access to your account, and for other purposes described in our privacy policy. 76284-692 76284-694. Techniques: Construct BsaI-HF®v2; Esp3I; BbsI-HF ® BsmBI-v2 ... BsaI-HFv2, BsmBI-v2 and T4 DNA Ligase are very stable, allowing cycling up to 60 cycles, with high efficiency and fidelity. Product Name: BsaI-HF®v2 Catalog Number: R3733L Concentration: 20,000 U/ml Unit Definition: One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 37°C in a total reaction volume of 50 µl. 5000 units ( 20,000 units/ml ) - Unavailable in your region: R3733S. Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. For difficult reaction, the s step time and number of cycles can be increased. BsaI-HF ® v2 10 U 0.5 MQ water 4 15 . NEB BsmBI-v2 NEB SpeI-HF NEB SalI-HF NEB PstI-HF NEB MluI-HF NEB BbsI-HF NEB AgeI HF NEB BsaI-HF®v2 NEB SNAP-Cell® Oregon Green® New England Biolabs (NEB) Gibson Assembly Cloning Kit NEB Gibson Assembly® Master Mix New England Biolabs (NEB) Deoxynucleotide Solution Mix New England Biolabs (NEB) Phusion High-Fidelity DNA Polymerase New England Biolabs (NEB) T7 … … High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. Place both PCR tube strips in a heat block set to 37°C. 5'GGTCTCN13'3'CCAGAGN55'Thermo Scientific Eco31I (BsaI) restriction enzyme recognizes GGTCTC(1/5)^ sites and cuts best at 37C in G buffer (Isoschizomers: BsaI, Bso31I, BspTNI). Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. NEBcutter™ v2.0 ; NEBioCalculator® REBASE® FAQs & Troubleshooting FAQs. This award was given to the company that judges felt had most impressed its customers. Set up 20 µl assembly reactions as follows: REAGENT ASSEMBLY REACTION; pGGAselect Destination Plasmid (1)* 1 μl: Amplicon Inserts (2) 3 nM (3) each DNA fragment (final concentration) T4 DNA Ligase Buffer (10X) 2 μl: NEB Golden Gate Enzyme Kit (BsaI-HFv2) 2 μl: Nuclease-free H 2 O : to 20 μl (4) *or user … Restriction Enzymes in Golden Gate Assembly Type IIS restriction enzymes have both recognition and binding sites, but cut downstream of the recognition site, creating 4-base overhangs … Code Name Size Quantity Price; R3733S. We regularly use 2 minute steps for the 37°C and 16°C steps with 20 cycles. Article Snippet: The reactions were cycled between 37°C and 16°C (SapI, Esp3I, BsaI-HFv2, BbsI-HF) or 42°C and 16°C (BsmBI-v2) for 5 minutes at each temperature for 30 cycles, and then subjected to a final heat-soak for 5 minutes at 60°C. Your credentials are incorrect or you are trying to login with a non-existing webshop account. HF enzymes also exhibit dramatically reduced star activity. NEB ® Golden Gate Assembly Kit (BsaI-HF ® v2) BsmBI-v2; Submit Restocking Order. Start a stop-watch to count up. 5000 units ( 20,000 units/ml ) - Unavailable in your region: R3733S. Insert assembly calls for careful design of overhangs to direct the assembly, as well as verification that the Type IIS REase sites used are not present in the fragments for the assembly of the expected product. BsaI-HF ® v2; T4 DNA Ligase; Esp3I; Submit Restocking Order. 1000 units-1 + Unavailable in your region. Catalog # Size Price (£) E1601L: 100 REACTIONS: £379.00: E1601S: 20 REACTIONS: £143.00: Learn about Ligase Fidelity and Push the Limits of Golden Gate Assembly (35+ fragments) Updated to include BsaI-HFv2 (optimized for Golden Gate) Seamless cloning - no scar remains following assembly Includes destination plasmid with … BsaI-HF®v2, New England Biolabs. BsmBI-v2; DNA Modifying Enzymes (Cloning) Immobilized T4 DNA Ligase; T5 Exonuclease; Tth Argonaute (TtAgo) NGS Sample Prep. Ineligible item added to cart. High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. NEB ® Golden Gate Assembly Kit (BsaI-HF ® v2) #E1601. 5000 units-1 + Unavailable in your region ®. What does HF® refer to following the name of a restriction enzyme? BsaI-HFv2. BsaI-HFv2 . This product has been discontinued and replaced by NEB ® Golden Gate Assembly Kit (BsaI-HF ® v2) (NEB #E1601) Catalog # E1600 was discontinued on October 18, 2018 ; Product Introduction. BsaI-HFv2. 1000 units-1 + Unavailable in your region. Product Name: NEB® Golden Gate Enzyme Kit (BsaI-HF®v2) Catalog Number: E1601L Packaging Lot Number: 10084258 Expiration Date: 03/2022 Storage Temperature: -20°C Specification Version: PS-E1601S/L v2.0 NEB® Golden Gate Enzyme Kit (BsaI-HF®v2) Component List NEB Part Number Component Description Lot Number Individual QC Result N0309AAVIAL pGGAselect Destination … Ineligible item added to cart. BsaI-HF ® v2 Product information. CiteAb Award "Researchers' Choice" Bethyl Laboratories Inc was honored with the 2021 CiteAb Award for "Researcher’s Choice". R3733L. ® ® ® ® ® ®. 24-Fragment Golden Gate Assembly using BsaI-HF ® v2 Overview. R3733L. Please review and update your order accordingly If you have any questions, please contact Customer Service at freezers@neb.com or 1-800-632-5227 x 8. HF enzymes also exhibit dramatically reduced star activity. BsaI-HF ® v2; Submit Restocking Order. Consider whether your workflow would be enhanced by adding more cycles. BsaI has a High Fidelity version BsaI-HF ® v2 . NEB ® Golden Gate Assembly Kit (BsaI-HF ® v2) Esp3I; BsaI-HF®v2; BbsI-HF ® Nuclease-free Water; Materials Sold Separately NEBuffer 3.1; Gel Loading Dye, Purple (6X), no SDS; FAQs Why is there a BsmBI site in LITMUS 38 but not LITMUS 39? Golden Gate Assembly of 24 fragments using BsaI-HFv2 restriction enzyme with T4 DNA Ligase and subsequent transformation into NEB 10-beta Competent E. coli (High Efficiency).. Materials 95% pure as determined by SDS-PAGE analysis using Coomassie Blue detection. Repeat step 4 with 12 µL of Diluent B and 4 µL of BsaI-HF v2 enzym e. 7. Which restriction enzymes are used in GoldenBraid Assembly? See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat i BsmBI-v2 NEB R0739 New England Biolabs. Protocol Golden Gate Assembly Protocol for Using NEB® Golden Gate Assembly Kit (BsaI-HF®v2) (E1601) Related products. Read more. Which restriction enzymes are used in Golden Gate Assembly? 1000 units ( 20,000 units/ml ) - Unavailable in your region: R3733L. When is star activity a … BsaI-HF®v2, New England Biolabs Supplier: New England Biolabs. BsaI-HFv2 . HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 minutes with the flexibility to … 1 hour and 45 minutes. BsaI has a High Fidelity version BsaI-HF ® v2 . Seamless cloning – no scar remains following assembly Includes destination plasmid with T7/SP6 promoters Ordered assembly of multiple fragments in a single reaction Can also be used for … BsaI-HF ® v2 Product information. High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. Do you need any help ordering? Our website shows product prices without login!Therefore webshop accounts that were only used to view prices have been deleted. NEBNext ® ARTIC SARS-CoV-2 Library Prep Kit (Illumina ®) NEBNext ® ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina ®) NEBNext ® ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies ® NEBNext ® Multiplex Oligos for Illumina ® (Unique … When is star activity a concern? 52-Fragment Golden Gate Assembly using BsaI-HF ® v2 (NEB #E1601) Videos . How does the level of star activity of BsaI-HF compare to BsaI? Customer care. For assembly, all the fragments were digested with BsaI-HF v2 (New England Biolabs) and purified directly using NucleoSpin Gel and PCR clean-up … Code Name Size Quantity Price; R3733S. BsaI has a High Fidelity version BsaI-HF ® v2 . Golden Gate reaction conditions: Golden Gate reaction conditions are flexible. 1000 units ( 20,000 units/ml ) - Unavailable in your region: R3733L. BsaI-HFv2. BsaI-HFv2.
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