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