Cite. 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. There was an issue creating your account. 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. Receive the latest news, hot plasmids, discounts and more. The insertion site is flanked by BstZI, EcoRI, and NotI sites. 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. Other amplification products including primer dimers will compete for ligation into the T vector, decreasing the possibility that the desired insert will be cloned. Please check your network settings and try again. Sign Up. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Vector Map (Click image to enlarge) × pGEM-T Vector Map. PCR cloning system for expression in mammalian cells. Home. There was an issue resetting your password. © 2021 GSL Biotech LLC | Sitemap | Privacy Policy | Legal Disclaimers. The insertion site is flanked by BstZI sites. Second-generation, high-performance GoTaq® G2 DNA Polymerase with Mg-free buffers. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Download SnapGene or SnapGene Viewer. Alternatively, T-Vector pMD20 retains the MCS of pUC19. Please try again or contact Customer Service. To minimize other competing products, gel purify the PCR fragment of interest. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. The pGEM-T Easy vector has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. The insertion site is flanked by BstZI sites. A password reset email has been sent to the primary email address associated with your account. Second-generation, high-performance GoTaq® G2 DNA Polymerase in a ready-to-use master mix. Please try again or contact Customer Service. A verified email address is required to access the full functionality of your Promega.com account. Subscribe to Our Blog. Please request another reset link. PGEM-T is a linearized cloning vector that can not be multiplied. Please update your browser to Internet Explorer 11 or above. 3rd Feb, 2016. Ready-to-use optimized master mix for room-temperature PCR assembly. TOP10, DH5α and TOP10F´, JM109. 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. T-Vector pMD19 (Simple) is derived from pUC19 and has deletions of all of the restriction enzyme sites in the multiple-cloning site (MCS). Sign Up for Our Newsletter. A verification email has been sent to the primary email address associated with your account. 製品マニュアル（日本語） DH5α使用説明書. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Introduction. Addgene is a nonprofit plasmid repository. When you select your country, you agree that we can place these functional cookies on your device. 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. Contact Us . The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. Multiple PCR products were amplified and cloned into the pGEM®-T or pGEM®-T Easy Vector. A3600. Have questions about your order, deposit, or a plasmid? To protect your privacy, your account has been locked after 6 failed login attempts. To protect your privacy, your account will be locked after 6 failed attempts. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Terms and Conditions The insertion site is flanked by BstZI, EcoRI, and NotI sites. There was an issue sending the verification email. There was an issue verifying your email address. Our records indicate that this email address is already registered. 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. Search our products with this vector backbone We offer a wide variety of inserts for this backbone. Please contact Customer Service to unlock your account. pGEM-T. Parental vector for TA cloning of PCR products. © 2021 Promega Corporation. This vector is also known as pGEM®‑5Zf(+). Legal and Trademarks Stay notified of Promega events, products and news. Is the insert size too big for a pGEM T easy vector? Your password reset link has expired. Unfortunately, I did not get any insert. Check your inbox to complete email verification. ACCESSION . Please try again or contact Customer Service. 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. We provide medical information and facilitate research collaborations. Parental vector for TA cloning of PCR products. The pGEM®-T Vector Systems are convenient for cloning PCR products. You have successfully reset your password. This vector is also known as pGEM®‑5Zf(+). 1 Recommendation. Subscribe. Most commercially available competent cells are appropriate for the plasmid, e.g. パフォーマンス. I am afraid you will have to buy it again and again. 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 pGEM®-T Vector System II 20 reactions A3610 Includes: • 1.2µg ®-T Vector (50ng/µl)pGEM • 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 × 200µl) PRODUCT SIZE CAT.# pGEM®-T Easy Vector System I 20 reactions A1360 Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. After that, you will need to contact Customer Service to unlock your account. Alternatively, a double digestion may be used to release the insert from the vector. The only difference between pGEM-T and pGEM-T Easy is in the multiple cloning site (MCS). 1. 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。 ベクターマップ＆シークエンス. Don't have either application? Please try again or contact Customer Service. Our customer and technical support experts are here to help! The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Contains GoTaq® G2 enzyme. The coding sequence was inserted by TA cloning. Thank you for verifying your email address. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. The pGEM®-T vectors are a popular choice for general PCR cloning. 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. There was an issue logging into your account. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. You've created a Promega.com account. Therefore, after cloning, restriction enzyme digestion analysis of the PCR insert is possible. Latest generation GoTaq® polymerase—high-performance for your everyday PCR needs. PCR cloning vectors with 3 options for insert excision. VERSION . Privacy Policy and Requests for Information Our website uses functional cookies that do not collect any personal information or track your browsing activity. The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. Introduction 1.A. Vector … The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. Let's find the product that meets your needs. To increase convenience, we tested conditions for shortening the ligation time. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. All Rights Reserved. Please try again or contact Customer Service. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. There was an issue with the password reset process. pGEM®-T Easy Parental vector for TA cloning of PCR products. The strand shcnvn is complementary to the ssDNA by this vector. Close. ×Please choose an application for opening sequence files. http://www.promega.com/products/pcr/pcr-cloning/pgem_t-easy-vector-systems/ : Video describing the use of the pGEM-T Vector Systems. pGEM®-T Vector System II 20 reactions A3610 For Laboratory Use. Estanis Navarro. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. This product is available through the Promega Helix onsite stocking program. To see this sequence with restriction sites, features, and translations, please download SnapGene or the free SnapGene Viewer. The T-overhangs at the insertion site greatly improve the efficiency of This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. Contact Addgene. 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. Please try again or contact Customer Service. A 15-minute ligation gave ~50% transformants by blue/white selection with further improvements when BSA was added. 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. X65308). This allows the insert DNA to be removed with a single restriction digest using either of these enzymes. 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. Search and compare our plasmid-based products. 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. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). LOCUS pGEM-T Easy 3015 bp ds-DNA circular SYN 25-NOV-2013 DEFINITION Parental vector for TA cloning of PCR products. Catalog number selected: Your commerce experience may be limited. 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. Congratulations! pGEM®-T Parental vector for TA cloning of PCR products. KEYWORDS pGEM-T Easy SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3015) AUTHORS Promega TITLE Direct Submission JOURNAL … There was an error processing your request. ベクターのT突出末端の安定性. クイックプロトコル (pGEM-T Vectors) 製品マニュアル. Learn about the latest plasmid technologies and research tools. Enter your username and we'll send a link to reset your password. We offer numerous convenient solutions to meet your lab's needs. I decided to use pGEM T easy for doing TA cloning before I obtain a properly digested PCR product. Note: You will not be able to access your account until your email is verified. 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. Map and Sequence File:    Download    Open. In addition, this vector contains the SP6 promoter upstream of MCS. See Protocol for detailed storage recommendations. Analyze Sequence: pGEM-T Easy Vector. Trademarks We've detected that you are using an older version of Internet Explorer. Home » Resources » Plasmid Files » Basic Cloning Vectors » pGEM-T Easy. Get in touch with a nearby distributor or sales representative. You have not verified your email address.
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