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