クイックプロトコル (pGEM-T Vectors) 製品マニュアル. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. & Engineering, Model Addgene is a nonprofit plasmid repository. 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. Introduction 1.A. PCR was employed to amplify cDNA fragments specific to TGF-α variant I, variant II, and wt TGF-α. パフォーマンス. The pGEM-T and pGEM-T Easy plasmid vectors are essentially the same but with one important difference. Test de protection de la ribonucléase. 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. Contact Addgene. PRINTED IN USA. ACCESSION . Linearize at EcoRV to create TA cloning vector. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Note: can —quencød using the fol- lowing pnmgs SP6 Promter Primer (Cat* aa)11) Promotg … The coding sequence was inserted by TA cloning. Subscribe to Our Blog. This website uses cookies to ensure you get the best experience. 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 . Revised 12/10 Part# TM042 The pGEM®-T Easy Vector Systems are convienent systems to clone PCR products. ベクターマップ＆シークエンス. Complete Protocol. Editing, Cloning The insertion site is flanked by BstZI, EcoRI, and NotI sites. X65308). The position of the T is indicated by * in the pGEM®-T Vector Sequence … pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. Full sequence for pGEM®-T Easy shared on Benchling. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. TOP10, DH5α and TOP10F´, JM109. produits PCR ont été intégrés dans le vecteur pGEM-T easy par clonage TA et ont ensuite été extraits par digestion. What strain of bacteria does my stab contain? If you run into any problems registering, depositing, or ordering please contact us at [email protected] Technical Manual pGEM®-T and pGEM®-T Easy Vector Systems INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380, A3600 AND A3610. 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. 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. 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. Don't have either application? 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. Have questions about your order, deposit, or a plasmid? What is virus associated DNA, and why do I have to order it? Systems, Research How do I place an order? Plusieurs tentatives d’insertion du p60 dans le vecteur navette ont été réalisées sans succès. Does Addgene accept orders by fax, phone or email? There is a problem with the plasmid I received. pGEM®-T Parental vector for TA cloning of PCR products. 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. Receive the latest news, hot plasmids, discounts and more. Revised 12/18 www.promega.com 1. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Most commercially available competent cells are appropriate for the plasmid, e.g. 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。 XX CC pGEM-T has dT, which improves efficiency of ligation of PCR product. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. 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. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Get … pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. 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. Download SnapGene or SnapGene Viewer. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. 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 … Copy Sequence. 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. The position of the T is indicated by * in the pGEM®-T Vector Sequence … Thus, several options exist to remove the desired insert DNA with a single restriction digestion. Sign Up for Our Newsletter. 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). The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. How can I be notified when a plasmid from a specific lab or paper is available? Learn about the latest plasmid technologies and research tools. ベクターのT突出末端の安定性. The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. Have questions about your order, deposit, or a plasmid? Analyze Sequence: pGEM-T Easy Vector. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. Procedure: 1. Quick Protocols. pGEM®-T Easy Vector System I 20 reactions A1360 For Laboratory Use. Contact Us . 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 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. Includes: 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. Protocols. KEYWORDS pGEM-T Easy SOURCE synthetic DNA construct … 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. Map and Sequence File:    Download    Open. 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. What do I need to know about the customs and importation process for my country? 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. © 2021 GSL Biotech LLC | Sitemap | Privacy Policy | Legal Disclaimers. The pGEM-T Easy vector has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. Name: pGEM-t-easy: Type: plasmid: Supplier: Description: The pGEM®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Determine the volume of PCR product to add to the ligation. Video Protocols. 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 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. PGem T easy suitable for insert Pcr product because it add poly a tail in pcr product. a. 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. Receive the latest news, hot plasmids, discounts and more. Bigger peace of pcr product hard to insert and ligase affect insertion. Do I need a new MTA for Penn viral vectors. Subscribe. Your professor will come around with the PGEM-T Easy Vector and T4 DNA ligase. This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. Specifications. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. Fields, Pathways By continuing to use this site, you agree to the use of cookies. Figure 3. pGEM®-T Easy Vector circle map and sequence reference points. Please note: Your browser does not support the features used on Addgene's website. VERSION . .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. X65308). Parental vector for TA cloning of PCR products. How can I track requests for my plasmids? 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. Genome & ORFs. Learn about the latest plasmid technologies and research tools. A chaque fois, les essais de transformation n’ont fourni que le pGEM-T easy religué. ×Please choose an application for opening sequence files. This vector is also known as pGEM®‑5Zf(+). Sign Up. Learn more, Please note: Your browser does not fully support some of the features used on Addgene's website. What is an MTA/Who is authorized to sign? The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. The insertion site is flanked by BstZI sites. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. 製品マニュアル（日本語） DH5α使用説明書. pGEM®-T Easy vector Sequence.