You have successfully reset your password. There was an issue with the password reset process. Please contact Customer Service to unlock your account. 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 Promega mission statement is: To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Our records indicate that this email address is already registered. A3600. All Rights Reserved. A verified email address is required to access the full functionality of your Promega.com account. Due to our annual Physical inventory orders received after 5PM Central Time on Tuesday March 9 will be shipped according to the following schedule: The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Using the pGEM-T-mH5 vector that we have previously ... Promega) was added and incubated for one additional hour. Legal and Trademarks X65308). 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. This product is available through the Promega Helix onsite stocking program. The insertion site is flanked by BstZI, EcoRI, and NotI sites. These samples were collected from Da Longshu village (a, 45 samples), Bai Shiyan village (b, 28 … Instructions for Use of Product(s) Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. There was an issue logging into your account. Please try again or contact Customer Service. Ligation Protocol 1. 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. Legal and Trademarks Instructions for Use of Product(s) A1360, A1380, A3600, A3610. Please contact Customer Service to unlock your account. Molecular Cloning: A Laboratory Manual Third Edition. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. 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. Background: Development of resistance to doxorubicin-based chemotherapy limits its curative effect in osteosarcoma. Please try again or contact Customer Service. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Canada orders: Ship Monday March 15 for arrival on Tuesday March 16. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. PLos ONE, Plate Readers, Fluorometers & Luminometers, Save 20% on pGL4 Luciferase Reporter Vectors, enter PGL20 at checkout. This page is informational only - this vector is NOT available from Addgene - please contact the manufacturer for further details. 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. Abstract. Stay notified of Promega events, products and news. The 12/18 version of this Technical Manual was revised to remove references to discontinued products in the notes on sequencing primers in Section 5.B. 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. Please request another reset link. Quick PROTOCOL 1 pGEM®-T and pGEM®-T Easy Vector Systems INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380, A3600 AND A3610. Terms and Conditions Your commerce experience may be limited. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). However, ratios of 8:1 to 1:8 have been used successfully. In this study, a specific 277-bp cDNA fragment of DHD4 was amplified and then cloned into the pGEM-T Easy vector (Promega), which was used to produce antisense and sense RNA probes. The green and blue arrows indicate 5′-RACE products characterized from WT and 1–910 EagI constructs. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Product Components and Storage Conditions PRODUCT SIZE CAT. ®Protocol for Ligations Using the pGEM -T and pGEM®-T Easy Vectors and the 2X Rapid Ligation Buffer 3.A. Plates were developed using 1 … The pGEM®-T and pGEM®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Issai Falcon. To protect your privacy, your account has been locked after 6 failed login attempts. Instructions for Use of Product(s) A1360, A1380, A3600, A3610. 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. You've created a Promega.com account. There was an issue sending the verification email. There was an issue logging into your account. The pGEM®-T and pGEM®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. When you select your country, you agree that we can place these functional cookies on your device. Description. Promega Notes 71 , 8–9. Download PDF. © 2021 Promega Corporation. # pGEM®-5Zf(+) Vector 20µg P2241 The pGEM®-5Zf(+) Vector is supplied with a glycerol stock of bacterial strain JM109. The assay sensitivity was determined using pGEM-T Easy Vector plasmids (Promega, Madison, WI) containing the target sequence of the N (961 bp) and S (1119 bp) genes of SARS-CoV-2. 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. US orders: Ship Saturday March 13 for arrival on Monday March 15. Please check your network settings and try again. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. PCR cloning system for expression in mammalian cells. 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. The gold arrows indicate 5′-RACE products characterized from the Δ5G construct. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. Please try again or contact Customer Service. When you select your country, you agree that we can place these functional cookies on your device. Literature # TM042. Page 4 Revised 5/07 GGAGA GCTCC CAACG CGTTG GATGC ATAGC TTGAG … 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. PCR products with low concentration or generating heterogeneity in the sequencing chromatograms were cloned into pGEM-T Easy Vector (Promega) for sequencing. There was an issue resetting your password. Complete Protocol The insertion site is flanked by BstZI sites. ... (T. aculeatus and three Zaglossus spp.) Molecular Cloning: A Laboratory Manual Third Edition, 1982. Second-generation, high-performance GoTaq® G2 DNA Polymerase with Mg-free buffers. 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. US orders: Ship Saturday March 13 for arrival on Monday March 15. Due to our annual Physical inventory orders received after 5PM Central Time on Tuesday March 9 will be shipped according to the following schedule: The shoot apex tissues of young seedlings were fixed in RNase-free formalin/acetic acid/alcohol fixative. We evaluated the cloning efficiency of different size PCR products into three T-vector cloning systems. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. We offer numerous convenient solutions to meet your lab's needs. Trademarks. 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. www.promega.com Part# TM042 Printed in USA. Revised 4/17 www.promega.com 2. A verification email has been sent to the primary email address associated with your account. Please try again or contact Customer Service. X65308). Diese Seite wurde zuletzt am … In the pGEM®-T Vector, T7 and SP6 RNA polymerase promoters flank a multiple cloning region within the α-peptide coding region for β-galactosidase. Second-generation, high-performance GoTaq® G2 DNA Polymerase in a ready-to-use master mix. The 3.9-kb product was cloned into pGEM-T Easy (Promega… Please check your network settings and try again. 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®-5Zf(+) Vector serves as a standard cloning vector and as a template for in vitro transcription. Enter your username and we'll send a link to reset your password. Note: You will not be able to access your account until your email is verified. 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. To protect your privacy, your account will be locked after 6 failed attempts. Get in touch with a nearby distributor or sales representative. 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. Frackman, S. and Kephart, D (1999) Rapid ligation for the pGEM®-T and pGEM®-T Easy Vector Systems. The pGEM®-T Vector cloning region is flanked by recognition sites for the enzyme BstZI. We've detected that you are using an older version of Internet Explorer. Ligation Using 2X Rapid Ligation Buffer 1. Privacy Policy and Requests for Information The high copy number pGEM®-T and pGEM®-T Easy Vectors contain T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the alpha-peptide coding region of the enzyme beta-galactosidase. PCR cloning vectors with 3 options for insert excision. This vector contains T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide coding region of β-galactosidase. The vector carries the lacZ alpha-peptide and the multiple cloning region arrangement from pUC18 allowing selection of recombinants by blue/white screening. 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. You have not verified your email address. Check your inbox to complete email verification. pGEM®-T Easy Parental vector for TA cloning of PCR products. The pGEM ®-T and pGEM -T Easy Vector Systems have been optimized using a 1:1 molar ratio of the Control Insert DNA to the vectors. Ratios from 3:1 to 1:3 provide good initial parameters. pGEM®-T Parental vector for TA cloning of PCR products. Introduction. You have not verified your email address. You have successfully reset your password. DNA concentration of linearised recombinant plasmid was determined using the Qubit 1× dsDNA HS Assay Kit (Invitrogen, CA, USA). 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. Download PDF. The pGEM®-T and pGEM®-T Easy Vector Systems gave a high number of recombinants across a broad range of insert sizes (0.5–3kb) while the TOPO TA Cloning® system worked well for inserts less than 1kb, but showed a striking decrease in performance with larger insert sizes (1–3kb). Catalog number selected: © 2021 Promega Corporation. A password reset email has been sent to the primary email address associated with your account. There was an error processing your request. 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 provide medical information and facilitate research collaborations. Summary of Changes Let's find the product that meets your needs. Our customer and technical support experts are here to help! The single major product was cloned into the pGEM-T-easy vector (Promega) and was sequenced. Reactions using this buffer may be incubated for 1 hour at room temperature. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. A1360, A1380, A3600, A3610. 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. However, the in vivo ECM is comprised not only of proteins but also of a variety of non-protein components. There was an issue creating your account. Thank you for verifying your email address. Enter your username and we'll send a link to reset your password. Our customer and technical support experts are here to help! There was an issue with the password reset process. Please try again or contact Customer Service. Shop Now ›, 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. Our website uses functional cookies that do not collect any personal information or track your browsing activity. This is a free resource for the scientific community that is compiled by Addgene.. Your password reset link has expired. Your password reset link has expired. Please try again or contact Customer Service. This paper. Briefly centrifuge the pGEM ®-T or pGEM -T Easy Vector and Control Insert DNA tubes to collect contents at the bottom of the tube. and Section 5.D. Download Full PDF Package. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. By creating an account, you confirm that you accept the, Plate Readers, Fluorometers & Luminometers, Privacy Policy and Requests for Information. A verified email address is required to access the full functionality of your Promega.com account. After that, you will need to contact Customer Service to unlock your account. After that, you will need to contact Customer Service to unlock your account. PDF (548k). Get in touch with a nearby distributor or sales representative. There was an issue sending the verification email. In addition, we excised the gene encoding GFP-mRNA-96-mer from pTRE-GFP-96-mer and inserted it into plasmid pGEM, because that plasmid contains a bacteriophage T7 promoter. Our records indicate that this email address is already registered. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. Alternatively, a double digestion may be used to release the insert from the vector. READ PAPER. Please try again or contact Customer Service. 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. In this study, we describe a method for producing armored L-RNA. However, the length of RNA packaged in the virus-like particles with high efficiency is usually less than 500 bases. Privacy Policy and Requests for Information 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 Main. Congratulations! 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. A short summary of this paper. Please try again or contact Customer Service. There was an issue creating your account. Contains GoTaq® G2 enzyme. We prepared a series of pGEM plasmids (Promega) containing 1, 2, 4, 8, 16, 32, or 64 tandem repeats of the sequence described above. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences.
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