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