We offer numerous convenient solutions to meet your lab's needs. Second-generation, high-performance GoTaq® G2 DNA Polymerase in a ready-to-use master mix. Trademarks The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. There was an issue verifying your email address. To protect your privacy, your account will be locked after 6 failed attempts. Receive the latest news, hot plasmids, discounts and more. pGEM®-T Easy Vector Systemは、従来のpGEM®-T Vector Systemの機能に加え、マルチクローニングサイトの両端にEcoRIとNotIサイトが加えられました。そのため、1種類（NotI、EcoRIあるいはBstZI）の制限酵素を用いるだけで、クローニング後のインサートDNAを簡単に切り出すことがきます。 The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. pGEM®-T Easy Parental vector for TA cloning of PCR products. We've detected that you are using an older version of Internet Explorer. The vectors are prepared by cutting the pGEM ®-5Zf(+) and pGEM ®-T Easy Vectors, respectively, with EcoR V and adding a 3´ terminal thymidine to both ends. pGEM-T. Parental vector for TA cloning of PCR products. I am afraid you will have to buy it again and again. Let's find the product that meets your needs. ベクターマップ＆シークエンス. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Home » Resources » Plasmid Files » Basic Cloning Vectors » pGEM-T Easy. The insertion site is flanked by BstZI sites. http://www.promega.com/products/pcr/pcr-cloning/pgem_t-easy-vector-systems/ : Video describing the use of the pGEM-T Vector Systems. Introduction 1.A. A verification email has been sent to the primary email address associated with your account. ACCESSION . Have questions about your order, deposit, or a plasmid? Second-generation, high-performance GoTaq® G2 DNA Polymerase with Mg-free buffers. This product is available through the Promega Helix onsite stocking program. Terms and Conditions 迅速なライゲーションバッファー添付によるキットの改良. A 15-minute ligation gave ~50% transformants by blue/white selection with further improvements when BSA was added. Thank you for verifying your email address. Ready-to-use optimized master mix for room-temperature PCR assembly. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Congratulations! Parental vector for TA cloning of PCR products. I decided to use pGEM T easy for doing TA cloning before I obtain a properly digested PCR product. Privacy Policy and Requests for Information The insertion site is flanked by BstZI sites. X65308). LOCUS pGEM-T Easy 3015 bp ds-DNA circular SYN 25-NOV-2013 DEFINITION Parental vector for TA cloning of PCR products. The pGEM®-T Vector was created by linearizing the pGEM®-5Zf(+) Vector with EcoRV at base 51 and adding a T to both 3´-ends. パフォーマンス. VERSION . Introduction. Our website uses functional cookies that do not collect any personal information or track your browsing activity. Estanis Navarro. We provide medical information and facilitate research collaborations. Home. Analyze Sequence: pGEM-T Easy Vector. In addition, this vector contains the SP6 promoter upstream of MCS. Multiple PCR products were amplified and cloned into the pGEM®-T or pGEM®-T Easy Vector. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. The pGEM€-T Vector has teen linearized with EcoR V at base SI of this sequence (indicata:l by an asterisk) and a T added to both 3 '-ends The added T is not included in this sequence The sequence shown comesponds to RNA synthesized by T7 RNA Polymerase and is complementary to RNA synthesized by SP6 RNA Polymerase. These single 3´-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmid by preventing recircularization of the vector and providing a compatible overhang for ligation of PCR products with A overhangs. There was an error processing your request. This vector is also known as pGEM®‑5Zf(+). Please try again or contact Customer Service. Please contact Customer Service to unlock your account. The strand shcnvn is complementary to the ssDNA by this vector. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Our customer and technical support experts are here to help! TOP10, DH5α and TOP10F´, JM109. PCR cloning system for expression in mammalian cells. Enter your username and we'll send a link to reset your password. A verified email address is required to access the full functionality of your Promega.com account. To see this sequence with restriction sites, features, and translations, please download SnapGene or the free SnapGene Viewer. You have not verified your email address. When you select your country, you agree that we can place these functional cookies on your device. PGEM-T is a linearized cloning vector that can not be multiplied. This allows the insert DNA to be removed with a single restriction digest using either of these enzymes. Note: You will not be able to access your account until your email is verified. Download SnapGene or SnapGene Viewer. Your password reset link has expired. The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. Legal and Trademarks © 2021 GSL Biotech LLC | Sitemap | Privacy Policy | Legal Disclaimers. ×Please choose an application for opening sequence files. Please try again or contact Customer Service. The MCS of the pGEM-T Easy Vector contains sequences on either side of the insert that are recognized by the restriction enzymes Not I and EcoR I. To protect your privacy, your account has been locked after 6 failed login attempts. Contact Us . 1. Latest generation GoTaq® polymerase—high-performance for your everyday PCR needs. A3600. The insertion site is flanked by BstZI, EcoRI, and NotI sites. Please try again or contact Customer Service. To increase convenience, we tested conditions for shortening the ligation time. Other amplification products including primer dimers will compete for ligation into the T vector, decreasing the possibility that the desired insert will be cloned. Cite. Check your inbox to complete email verification. Please check your network settings and try again. See Protocol for detailed storage recommendations. © 2021 Promega Corporation. Please try again or contact Customer Service. One of the easiest methods for cloning blunt-ended DNA fragments including PCR products is T-vector cloning, such as with pGEM®-T or pGEM®-T Easy Vector Systems.This method takes advantage of the “A” overhang added by a PCR enzyme like Taq DNA Polymerase.T vectors are linearized plasmids that have been treated to add 3′ T overhangs to match the A overhangs of the insert. クイックプロトコル (pGEM-T Vectors) 製品マニュアル. Contact Addgene. There was an issue with the password reset process. Please update your browser to Internet Explorer 11 or above. The insertion site is flanked by BstZI, EcoRI, and NotI sites. There was an issue resetting your password. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. The pGEM®-T vectors are a popular choice for general PCR cloning. Addgene is a nonprofit plasmid repository. These vectors are ready to use in ligation reactions; prepared by cutting with a restriction endonuclease that creates a blunt end and adding a 3´ terminal thymidine (T) to both ends. KEYWORDS pGEM-T Easy SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3015) AUTHORS Promega TITLE Direct Submission JOURNAL … These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Close. Contains GoTaq® G2 enzyme. PCR cloning vectors with 3 options for insert excision. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Our records indicate that this email address is already registered. ベクターのT突出末端の安定性. Please try again or contact Customer Service. The pGEM-T Easy vector has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. After that, you will need to contact Customer Service to unlock your account. The only difference between pGEM-T and pGEM-T Easy is in the multiple cloning site (MCS). To minimize other competing products, gel purify the PCR fragment of interest. This vector is also known as pGEM®‑5Zf(+). Don't have either application? Learn about the latest plasmid technologies and research tools. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. Your commerce experience may be limited. Unfortunately, I did not get any insert. In the pGEM®-T Vector, T7 and SP6 RNA polymerase promoters flank a multiple cloning region within the α-peptide coding region for β-galactosidase. Sign Up. The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. Search our products with this vector backbone We offer a wide variety of inserts for this backbone. 製品マニュアル（日本語） DH5α使用説明書. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. There was an issue sending the verification email. You've created a Promega.com account. Vector Features T-Overhangs for Easy PCR Cloning: The pGEM®-T and pGEM®-T Easy Vectors(a,b) are linearized vectors with a single 3´-terminal thymidine at both ends. 3rd Feb, 2016. Most commercially available competent cells are appropriate for the plasmid, e.g. The pGEM®-T Vector Systems are convenient for cloning PCR products. Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. Catalog number selected: pGEM®-T Parental vector for TA cloning of PCR products. Stay notified of Promega events, products and news. Alternatively, T-Vector pMD20 retains the MCS of pUC19. pGEM®-T Easy Vector System II 20 reactions A1380 Includes: • 1.2µg pGEM ®-T Easy Vector (50ng/µl) • 12µl Control Insert DNA (4ng/µl) • 100u T4 DNA Ligase • 200µl 2X Rapid Ligation Buffer, T4 DNA Ligase • 1.2ml JM109 Competent Cells, High Efficiency (6 x 200µl) • 1 Protocol Storage Conditions: Store the Competent Cells at –70°C. PLos ONE, Plate Readers, Fluorometers & Luminometers, Rapid Ligation for the pGEM®-T and pGEM®-T Easy Vector Systems, Comparing Cloning Efficiency of the pGEM®-T and pGEM®-T Easy Vectors to the TOPO TA Cloning® Vectors, Shorten the Ligation Time for the pGEM®-T Vector Systems, TRE5-A retrotransposition profiling reveals putative RNA polymerase III transcription complex binding sites on the, Privacy Policy and Requests for Information, Insert excision with a BstZI single digest, Ligation can be completed in 1 hour at room temperature, Available with or without competent cells. By creating an account, you confirm that you accept the, pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual, pGEM T and pGEM T Easy Vector Systems FB033, 2017 Get in touch with a nearby distributor or sales representative. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Subscribe. A password reset email has been sent to the primary email address associated with your account. Search and compare our plasmid-based products. The pGEM®-T Vector is ready to use in ligation reactions, prepared by cutting the pGEM®-5Zf(+) Vector with EcoRV and adding a 3´ terminal thymidine to both ends. The vectors are prepared by cutting the pGEM ®-5Zf(+) and pGEM ®-T Easy Vectors, respectively, with EcoR V and adding a 3´ terminal thymidine to both ends. You have successfully reset your password. Please request another reset link. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Map and Sequence File:    Download    Open. pGEM®-T Vector System II 20 reactions A3610 For Laboratory Use. There was an issue creating your account. Vector Map (Click image to enlarge) × pGEM-T Vector Map. Subscribe to Our Blog. All Rights Reserved. Vector … There was an issue logging into your account. The T-overhangs at the insertion site greatly improve the efficiency of Is the insert size too big for a pGEM T easy vector? Therefore, after cloning, restriction enzyme digestion analysis of the PCR insert is possible. T-Vector pMD19 (Simple) is derived from pUC19 and has deletions of all of the restriction enzyme sites in the multiple-cloning site (MCS).

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