Please try again or contact Customer Service. Ready-to-use optimized master mix for room-temperature PCR assembly. Enter your username and we'll send a link to reset your password. Promega Notes 71 , 8–9. When you select your country, you agree that we can place these functional cookies on your device. Terms and Conditions The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Please request another reset link. 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. There was an issue creating your account. Instructions for Use of Product(s) A1360, A1380, A3600, A3610. A short summary of this paper. The vector carries the lacZ alpha-peptide and the multiple cloning region arrangement from pUC18 allowing selection of recombinants by blue/white screening. READ PAPER. Diese Seite wurde zuletzt am … Our website uses functional cookies that do not collect any personal information or track your browsing activity. The pGEM®-3Zf(+) and pGEM®-3Zf(–) Vectors contain T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide coding region of β-galactosidase. Please try again or contact Customer Service. This vector is also known as pGEM®‑5Zf(+). Our website uses functional cookies that do not collect any personal information or track your browsing activity. Your password reset link has expired. Stay notified of Promega events, products and news. A verification email has been sent to the primary email address associated with your account. pGEM®-T Parental vector for TA cloning of PCR products. The pGEM-3Z Vector is intended for use as a standard cloning vector, as well as for the highly efficient synthesis of RNA in vitro. Due to our annual Physical inventory orders received after 5PM Central Time on Tuesday March 9 will be shipped according to the following schedule: and Section 5.D. The pGEM®-T and pGEM®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. To protect your privacy, your account will be locked after 6 failed attempts. We have successfully cloned PCR products generated using GoTaq® and GoTaq® Flexi DNA Polymerases into the pGEM®-T (Cat.# A3600), pGEM®-T Easy (Cat.# A1360) and pTARGET™ (Cat.# A1410) Vectors. Due to our annual Physical inventory orders received after 5PM Central Time on Tuesday March 9 will be shipped according to the following schedule: Revised 4/17 www.promega.com 2. However, ratios of 8:1 to 1:8 have been used successfully. To protect your privacy, your account will be locked after 6 failed attempts. Our customer and technical support experts are here to help! Download Full PDF Package. Check your inbox to complete email verification. Let's find the product that meets your needs. The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. There was an error processing your request. After that, you will need to contact Customer Service to unlock your account. Abstract. Please try again or contact Customer Service. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. Thank you for verifying your email address. There was an issue verifying your email address. DNA concentration of linearised recombinant plasmid was determined using the Qubit 1× dsDNA HS Assay Kit (Invitrogen, CA, USA). All Rights Reserved. Legal and Trademarks Download Free PDF. PCR products were gel-purified, cloned into the pGEM-T Easy Vector system (Promega Corporation, WI, USA) and sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems). Stay notified of Promega events, products and news. Ligation Using 2X Rapid Ligation Buffer 1. In this study, we describe a method for producing armored L-RNA. Please check your network settings and try again. Download PDF. RNase-resistant, noninfectious virus-like particles containing exogenous RNA sequences (armored RNA) are good candidates as RNA controls and standards in RNA virus detection. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. Page 4 Revised 5/07 GGAGA GCTCC CAACG CGTTG GATGC ATAGC TTGAG … Second-generation, high-performance GoTaq® G2 DNA Polymerase in a ready-to-use master mix. Ratios from 3:1 to 1:3 provide good initial parameters. In the current study, we focused on investigating the mechanisms underlying the development of doxorubicin resistance in osteosarcoma.Methods: The human osteosarcoma cell line MG-63 and doxorubicin-resistant MG-63/Dox cells were used in this study. Introduction. Insertional inactivation of the alpha-peptide allows recombinant clones to be directly identified by blue/white screening on indicator plates. You have successfully reset your password. Please try again or contact Customer Service. Terms and Conditions ®Briefly centrifuge the pGEM-T or pGEM®-T Easy Vector and Control Insert … A password reset email has been sent to the primary email address associated with your account. Get in touch with a nearby distributor or sales representative. Your commerce experience may be limited. We offer numerous convenient solutions to meet your lab's needs. Legal and Trademarks 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. As a result, PCR products amplified using GoTaq® DNA Polymerase will contain A-overhangs which makes it suitable for T-vector cloning. Plates were developed using 1 … The pGEM®-T and pGEM®-T Easy Vector Systems are convenient systems for the cloning of PCR products. A1360, A1380, A3600, A3610. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products.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. Our records indicate that this email address is already registered. ®Protocol for Ligations Using the pGEM -T and pGEM®-T Easy Vectors and the 2X Rapid Ligation Buffer 3.A. Our customer and technical support experts are here to help! These single 3´-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmids by preventing recircularization of the vector and providing a compatible overhang for PCR products generated by certain thermostable polymerases. These samples were collected from Da Longshu village (a, 45 samples), Bai Shiyan village (b, 28 … The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Please try again or contact Customer Service. The insertion site is flanked by BstZI, EcoRI, and NotI sites. PDF (548k). The pGEM®-11Zf(+) Vector is a standard cloning vector that contains T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide coding region of β-galactosidase. PCR cloning vectors with 3 options for insert excision. Issai Falcon. The pGEM®-3Z Vector is intended for use as a standard cloning vector, as well as for the highly efficient synthesis of RNA in vitro. We prepared a series of pGEM plasmids (Promega) containing 1, 2, 4, 8, 16, 32, or 64 tandem repeats of the sequence described above. There was an issue resetting your password. Product Components and Storage Conditions PRODUCT SIZE CAT.# pGEM®-7Zf(+) Vector 20µg P2251 The pGEM®-7Zf(+) Vector is provided with a glycerol stock of bacterial strain JM109. 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. Accordingly, as a means of enhancing tissue invasion, tumor cells use matrix metalloproteinases to degrade ECM proteins. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. There was an issue creating your account. X65308). Pod pepper (Capsicum frutescens) is widely planted in China, especially around Wenshan city, Yunnan province, and viral diseases have now also become a major threat to pepper production in Yunnan.During July 2019, 89 pepper leaf samples were collected from three different fields in Wenshan. The pGEM®-T Easy Vector multiple cloning region is flanked by recognition sites for the restriction enzymes EcoRI, BstZI and NotI, providing three single-enzyme digestions for release of the insert. To protect your privacy, your account has been locked after 6 failed login attempts. You have not verified your email address. However, the in vivo ECM is comprised not only of proteins but also of a variety of non-protein components. You have successfully reset your password. Instructions for Use of Product(s) A1360, A1380, A3600, A3610. Congratulations! Congratulations! 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. Using the pGEM-T-mH5 vector that we have previously ... Promega) was added and incubated for one additional hour. 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. Please try again or contact Customer Service. The 3.9-kb product was cloned into pGEM-T Easy (Promega… Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. We provide medical information and facilitate research collaborations. Complete Protocol The positive samples in this study were termed using the abbreviated name … Thus, several options exist to remove the desired insert DNA with a single restriction digestion. Revised 12/18 www.promega.com 3. There was an issue verifying your email address. Ligation Protocol 1. Die In-vitro-Transkription, also die DNA-abhängige Synthese von RNA im Reagenzglas, ist eine molekularbiologische Methode zur Erzeugung von RNA und zur Untersuchung von Promotoren und ihrer Aktivierung durch Transkriptionsfaktoren. This product is available through the Promega Helix onsite stocking program. Privacy Policy and Requests for Information These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. There was an error processing your request. US orders: Ship Saturday March 13 for arrival on Monday March 15. Dismiss. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. Reactions using this buffer may be incubated for 1 hour at room temperature. A password reset email has been sent to the primary email address associated with your account. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products.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 PDF. Trademarks. 36 Full PDFs related to this paper. We provide medical information and facilitate research collaborations. Main. This paper. Please update your browser to Internet Explorer 11 or above. Please try again or contact Customer Service. 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. Frackman, S. and Kephart, D (1999) Rapid ligation for the pGEM®-T and pGEM®-T Easy Vector Systems. A verified email address is required to access the full functionality of your Promega.com account. We evaluated the cloning efficiency of different size PCR products into three T-vector cloning systems. Privacy Policy and Requests for Information Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. There was an issue sending the verification email. Molecular Cloning: A Laboratory Manual Third Edition. 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. However, the length of RNA packaged in the virus-like particles with high efficiency is usually less than 500 bases. The insertion site is flanked by BstZI sites. The green and blue arrows indicate 5′-RACE products characterized from WT and 1–910 EagI constructs. There was an issue with the password reset process. PLos ONE, Plate Readers, Fluorometers & Luminometers, Save 20% on pGL4 Luciferase Reporter Vectors, enter PGL20 at checkout. Thank you for verifying your email address. The pGEM-T Vector is prepared by cutting Promega's pGEM-5Zf(+) Vector with EcoR V and adding a 3' terminal thymidine to both ends.These single 3'-T overhangs at the insertion site greatly improve the efficiency of ligation of a PCR product into the plasmid. This is a free resource for the scientific community that is compiled by Addgene.. There was an issue with the password reset process. A verification email has been sent to the primary email address associated with your account. To protect your privacy, your account has been locked after 6 failed login attempts. 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. After that, you will need to contact Customer Service to unlock your account. Download PDF. pGEM®-T Easy Parental vector for TA cloning of PCR products. https://doi.org/10.1530/JME-17-0142 http://jme.endocrinology-journals.org 2018 Society for Endocrinology Printed in Great Britain Published by Bioscientifica Ltd. Please contact Customer Service to unlock your account. A verified email address is required to access the full functionality of your Promega.com account. 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. Please check your network settings and try again. All Rights Reserved. Please request another reset link. Please try again or contact Customer Service.