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

Compte Fortnite Switch Gratuit, Série Extrême Limite Lieu Tournage, Brevet 2021 Annulé Covid, L'invention De L'écriture Ce2 Lutin Bazar, Quizz Musical 6 8 Ans, Les Brigades Du Tigre Film Youtube, Dieu Rend Justice, Vw Golf 2 Gti à Vendre,