How can I be notified when a plasmid from a specific lab or paper is available? The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. X65308). Don't have either application? Name: pGEM-t-easy: Type: plasmid: Supplier: Description: The pGEM®-T Easy Vector Systems are convenient systems for the cloning of PCR products. The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. Subscribe to Our Blog. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. PGem T easy suitable for insert Pcr product because it add poly a tail in pcr product. Home » Resources » Plasmid Files » Basic Cloning Vectors » pGEM-T. To see this sequence with restriction sites, features, and translations, please download SnapGene or the free SnapGene Viewer. Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. Copy Sequence. The pGEM-T and pGEM-T Easy plasmid vectors are essentially the same but with one important difference. How do I place an order? The pGEM®-T Easy Vector Systems are convienent systems to clone PCR products. How can I track requests for my plasmids? Quick Protocols. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Sign Up. What do I need to know about the customs and importation process for my country? The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. PRINTED IN USA. Parental vector for TA cloning of PCR products. Download SnapGene Map(.dna) Download GeneBank File(.gb) LOCUS Exported 3015 bp ds-DNA circular SYN 25-11-2013 DEFINITION Parental vector for TA cloning of PCR products. ACCESSION . ×Please choose an application for opening sequence files. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. The pGEM®-T Easy Vector Systems offer all of the advantages of the pGEM®-T Vector Systems with the added convenience of recognition sites for BstZI, EcoRI and NotI flanking the insertion site. Linearize at EcoRV to create TA cloning vector. 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 Product Size Cat.# pGEM®-T Easy Vector System II 20 reactions A1380 For Laboratory Use. By continuing to use this site, you agree to the use of cookies. Have questions about your order, deposit, or a plasmid? pGEM-T Sequencing Primers M13 Forward Sequence - 5’-CACGACGTTGTAAAACGAC-3’ M13 Reverse Sequence - 5’-GGATAACAATTTCACACAGG-3’ Sequencing Ambiguities R = A or G Y= C or T M= A or C K= G or T S= G or C W= A or T H= A, T or C B= G, T or C D= G, A or T V= G, A or C N= G, A, T or C . pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. 製品マニュアル（日本語） DH5α使用説明書. Protocols. This vector is also known as pGEM®‑5Zf(+). The insertion site is flanked by BstZI sites. Plusieurs tentatives d’insertion du p60 dans le vecteur navette ont été réalisées sans succès. pGEM®-T Easy Vector Sequence reference points: T7 RNA Polymerase transcription initiation site 1 SP6 RNA Polymerase transcription initiation site 141 T7 RNA Polymerase promoter 3002-6 SP6 RNA Polymerase promoter 136-158 multiple cloning site 10-128 lacZ start codon 180 lacoperon sequences 2839-2999, 166-395 lacoperator 100-216 β-lactamase coding region 1337-2197 phage f1 region 2383 … Specifications. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. pGEM®-T Parental vector for TA cloning of PCR products. XX CC pGEM-T has dT, which improves efficiency of ligation of PCR product. Receive the latest news, hot plasmids, discounts and more. The Promega mission statement is: To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Full sequence for pGEM®-T Easy shared on Benchling. Get … Determine the volume of PCR product to add to the ligation. ベクターのT突出末端の安定性. & Engineering, Model 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 parent vector is linearized at the position indicated by * in this pGEM®-T Easy Vector Sequence and a "T" is added at each end. Analyze Sequence: pGEM-T Easy Vector. If you run into any problems registering, depositing, or ordering please contact us at [email protected] There is a problem with the plasmid I received. PCR was employed to amplify cDNA fragments specific to TGF-α variant I, variant II, and wt TGF-α. Do I need a new MTA for Penn viral vectors. Technical Manual pGEM®-T and pGEM®-T Easy Vector Systems INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380, A3600 AND A3610. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. The pGEM-T Easy vector has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. Download SnapGene or SnapGene Viewer. Editing, Cloning Learn about the latest plasmid technologies and research tools. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. クイックプロトコル (pGEM-T Vectors) 製品マニュアル. Video Protocols. pGEM®-T Easy Vector System I 20 reactions A1360 For Laboratory Use. Your professor will come around with the PGEM-T Easy Vector and T4 DNA ligase. What is an MTA/Who is authorized to sign? 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. Systems, Research Please note: Your browser does not support the features used on Addgene's website. In this study the performance of the 2X Rapid Ligation Buffer is compared with that of the previously supplied T4 DNA Ligase 10X Buffer in both one-hour and 16-hour ligation reactions. Sign Up for Our Newsletter. TOP10, DH5α and TOP10F´, JM109. Genome The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. © 2021 GSL Biotech LLC | Sitemap | Privacy Policy | Legal Disclaimers. a. & ORFs. Figure 3. pGEM®-T Easy Vector circle map and sequence reference points. The Promega mission statement is: To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Procedure: 1. 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. Most commercially available competent cells are appropriate for the plasmid, e.g. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. What is virus associated DNA, and why do I have to order it? The coding sequence was inserted by TA cloning. produits PCR ont été intégrés dans le vecteur pGEM-T easy par clonage TA et ont ensuite été extraits par digestion. Revised 12/10 Part# TM042 pGEM®-T Easy vector Sequence. Have questions about your order, deposit, or a plasmid? Test de protection de la ribonucléase. Fields, Pathways Note: can —quencød using the fol- lowing pnmgs SP6 Promter Primer (Cat* aa)11) Promotg … VERSION . Thus, several options exist to remove the desired insert DNA with a single restriction digestion. パフォーマンス. Includes: Does Addgene accept orders by fax, phone or email? The position of the T is indicated by * in the pGEM®-T Vector Sequence … 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 insertion site is flanked by BstZI, EcoRI, and NotI sites. This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. REQUIRED MATERIALS PGEM-T Easy plasmid (Kit ordered from Fisher PR-1380) 2x rapid ligation buffer T4 DNA Ligase enzyme **Note a few ingredients to the Ligation reaction are NOT on your desk due to the very small volumes needed. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Complete Protocol. .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. 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. Learn about the latest plasmid technologies and research tools. Map and Sequence File:    Download    Open. Subscribe. What strain of bacteria does my stab contain? Contact Us . Addgene is a nonprofit plasmid repository. KEYWORDS pGEM-T Easy SOURCE synthetic DNA construct … ベクターマップ＆シークエンス. Receive the latest news, hot plasmids, discounts and more. The pGEM®-T and pGEM®-T Easy Systems are now provided with a new 2X Rapid Ligation Buffer that allows the user to perform ligation reactions in as little as one hour. X65308). A chaque fois, les essais de transformation n’ont fourni que le pGEM-T easy religué. They offer all of the advantages of the pGEM®-T Vector Systems with the added convenience of recognition sites for EcoRI and NotI flanking the insertion site. 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. They offer all of the advantages of the pGEM®-T Vector Systems with added convieneice of recognition sites for EcoRI and NotI flankin the insertion site. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Vector Features T-Overhangs for Easy PCR Cloning: The pGEM ®-T and pGEM -T Easy Vectors are linearized vectors with a single 3´-terminal thymidine at both ends. Introduction 1.A. Le produit de PCR a été clone dans le vecteur pGEM-T Easy (Promega) et séquence en utilisant des amorces imbriquées et la séquenase T7 (Amersham). .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. 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. Learn more, Download our file to copy and paste plasmid data, Open collection of AAV data generously shared by scientists, Basic analysis for a user-entered sequence; includes restriction sites and map, Digital collection of empty plasmid backbones from publications and commercially available sources. The position of the T is indicated by * in the pGEM®-T Vector Sequence … This website uses cookies to ensure you get the best experience. Revised 12/18 www.promega.com 1. Contact Addgene. Bigger peace of pcr product hard to insert and ligase affect insertion.