# pGEM®-5Zf(+) Vector 20µg P2241 The pGEM®-5Zf(+) Vector is supplied with a glycerol stock of bacterial strain JM109. Enter your username and we'll send a link to reset your password. Please try again or contact Customer Service. There was an issue logging into your account. A short summary of this paper. Please try again or contact Customer Service. Dismiss. 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. This paper. You've created a Promega.com account. Please try again or contact Customer Service. Latest generation GoTaq® polymerase—high-performance for your everyday PCR needs. 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. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. We prepared a series of pGEM plasmids (Promega) containing 1, 2, 4, 8, 16, 32, or 64 tandem repeats of the sequence described above. 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. There was an issue logging into your account. Our customer and technical support experts are here to help! 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. Second-generation, high-performance GoTaq® G2 DNA Polymerase in a ready-to-use master mix. Trademarks. Diese Seite wurde zuletzt am … Molecular Cloning: A Laboratory Manual Third Edition. 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. You've created a Promega.com account. 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. A verification email has been sent to the primary email address associated with your account. Our records indicate that this email address is already registered. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. 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. US orders: Ship Saturday March 13 for arrival on Monday March 15. The incubation period may be extended to increase the number of colonies after transformation. Please contact Customer Service to unlock your account. The insertion site is flanked by BstZI, EcoRI, and NotI sites. 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. You have not verified your email address. This vector contains T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide coding region of β-galactosidase. As a result, PCR products amplified using GoTaq® DNA Polymerase will contain A-overhangs which makes it suitable for T-vector cloning. Main. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Get in touch with a nearby distributor or sales representative. The pGEM®-T and pGEM®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. 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. 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. © 2021 Promega Corporation. 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. ... (T. aculeatus and three Zaglossus spp.) (2007) A new 10-min ligation method using a modified buffer system with a very low amount of T4 DNA ligase: the "coffee break ligation" technique. Issai Falcon. You have successfully reset your password. Accordingly, as a means of enhancing tissue invasion, tumor cells use matrix metalloproteinases to degrade ECM proteins. Note: You will not be able to access your account until your email is verified. Introduction. There was an issue with the password reset process. 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 website uses functional cookies that do not collect any personal information or track your browsing activity. There was an issue verifying your email address. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Terms and Conditions Let's find the product that meets your needs. and Section 5.D. See Protocol for detailed storage recommendations. However, ratios of 8:1 to 1:8 have been used successfully. Welcome to Vector Database!. Stay notified of Promega events, products and news. Background: Development of resistance to doxorubicin-based chemotherapy limits its curative effect in osteosarcoma. There was an issue with the password reset process. 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. 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. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. Abstract. Revised 4/17 www.promega.com 2. There was an issue creating your account. By creating an account, you confirm that you accept the, Plate Readers, Fluorometers & Luminometers, Privacy Policy and Requests for Information. DNA concentration of linearised recombinant plasmid was determined using the Qubit 1× dsDNA HS Assay Kit (Invitrogen, CA, USA). 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. PLos ONE, Plate Readers, Fluorometers & Luminometers, Save 20% on pGL4 Luciferase Reporter Vectors, enter PGL20 at checkout. A verified email address is required to access the full functionality of your Promega.com account. The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. To protect your privacy, your account has been locked after 6 failed login attempts. Please try again or contact Customer Service. 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. Check your inbox to complete email verification. The vector carries the lacZ alpha-peptide and the multiple cloning region arrangement from pUC18 allowing selection of recombinants by blue/white screening. We offer numerous convenient solutions to meet your lab's needs. 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. There was an issue resetting your password. Legal and Trademarks The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. Please try again or contact Customer Service. Download Free PDF. 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. 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. After that, you will need to contact Customer Service to unlock your account. 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 Literature # TM042. There was an error processing your request. Our website uses functional cookies that do not collect any personal information or track your browsing activity. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. The positive samples in this study were termed using the abbreviated name … Briefly centrifuge the pGEM ®-T or pGEM -T Easy Vector and Control Insert DNA tubes to collect contents at the bottom of the tube. Download PDF. There was an issue verifying your email address. The pGEM®-T Vector cloning region is flanked by recognition sites for the enzyme BstZI. The pGEM®-T and pGEM®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Using the pGEM-T-mH5 vector that we have previously ... Promega) was added and incubated for one additional hour. We've detected that you are using an older version of Internet Explorer. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. 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. PDF (548k). Instructions for Use of Product(s) A1360, A1380, A3600, A3610. The 3.9-kb product was cloned into pGEM-T Easy (Promega… Frackman, S. and Kephart, D (1999) Rapid ligation for the pGEM®-T and pGEM®-T Easy Vector Systems. To protect your privacy, your account has been locked after 6 failed login 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. 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. Revised 12/18 www.promega.com 3. The insertion site is flanked by BstZI sites. .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. Your password reset link has expired. Promega Notes 71 , 8–9. Due to our annual Physical inventory orders received after 5PM Central Time on Tuesday March 9 will be shipped according to the following schedule: Please request another reset link. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. Insertional inactivation of the alpha-peptide allows recombinant clones to be directly identified by blue/white screening on indicator plates. Please check your network settings and try again. Check your inbox to complete email verification. ®Briefly centrifuge the pGEM-T or pGEM®-T Easy Vector and Control Insert … 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. Canada orders: Ship Monday March 15 for arrival on Tuesday March 16. Summary of Changes 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. Canada orders: Ship Monday March 15 for arrival on Tuesday March 16. Please try again or contact Customer Service. Enter your username and we'll send a link to reset your password. 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. 36 Full PDFs related to this paper. Your password reset link has expired. Ratios from 3:1 to 1:3 provide good initial parameters. There was an issue sending the verification email. In this study, we describe a method for producing armored L-RNA. To protect your privacy, your account will be locked after 6 failed attempts. However, the in vivo ECM is comprised not only of proteins but also of a variety of non-protein components. The shoot apex tissues of young seedlings were fixed in RNase-free formalin/acetic acid/alcohol fixative. Plates were developed using 1 … 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. The single major product was cloned into the pGEM-T-easy vector (Promega) and was sequenced. This is a free resource for the scientific community that is compiled by Addgene.. 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. Get in touch with a nearby distributor or sales representative. 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. Alternatively, a double digestion may be used to release the insert from the vector. Please try again or contact Customer Service. A1360, A1380, A3600, A3610. The extracellular matrix (ECM) plays an important role in maintaining tissue homeostasis and poses a significant physical barrier to in vivo cell migration. When you select your country, you agree that we can place these functional cookies on your device. US orders: Ship Saturday March 13 for arrival on Monday March 15. Yoshino, Y., Ishida, M. and Horii, A. PCR products with low concentration or generating heterogeneity in the sequencing chromatograms were cloned into pGEM-T Easy Vector (Promega) for sequencing. We provide medical information and facilitate research collaborations. A password reset email has been sent to the primary email address associated with your account. There was an issue resetting your password. Download Full PDF Package. pGEM®-T Parental vector for TA cloning of PCR products. To protect your privacy, your account will be locked after 6 failed attempts. Ligation Using 2X Rapid Ligation Buffer 1. There was an issue creating your account. Trademarks. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. All Rights Reserved. Privacy Policy and Requests for Information The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Instructions for Use of Product(s) X65308). There was an error processing your request. These samples were collected from Da Longshu village (a, 45 samples), Bai Shiyan village (b, 28 … Stay notified of Promega events, products and news. Page 4 Revised 5/07 GGAGA GCTCC CAACG CGTTG GATGC ATAGC TTGAG … Congratulations! pGEM®-T Easy Parental vector for TA cloning of PCR products. Promega Corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, high-value products for the life sciences. 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. Your commerce experience may be limited. After that, you will need to contact Customer Service to unlock your account. Download PDF. Quick PROTOCOL 1 pGEM®-T and pGEM®-T Easy Vector Systems INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380, A3600 AND A3610. Revised 4/17 www.promega.com 2. Literature # TM042. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Catalog number selected: 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. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. 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. 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. 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. This page is informational only - this vector is NOT available from Addgene - please contact the manufacturer for further details. Ligation Protocol 1. Thank you for verifying your email address. All Rights Reserved. We evaluated the cloning efficiency of different size PCR products into three T-vector cloning systems. However, ratios of 8:1 to 1:8 have been used successfully. Reactions using this buffer may be incubated for 1 hour at room temperature. 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. 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. Complete Protocol Privacy Policy and Requests for Information 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. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. 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。 Legal and Trademarks 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. PCR cloning system for expression in mammalian cells. This product is available through the Promega Helix onsite stocking program. You have successfully reset your password. Ratios from 3:1 to 1:3 provide good initial parameters. Molecular Cloning: A Laboratory Manual Third Edition, 1982. A3600. 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. Please try again or contact Customer Service. PCR cloning vectors with 3 options for insert excision. When you select your country, you agree that we can place these functional cookies on your device. Thank you for verifying your email address. Download PDF. This vector is also known as pGEM®‑5Zf(+). Description. However, the length of RNA packaged in the virus-like particles with high efficiency is usually less than 500 bases. There was an issue sending the verification email. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Our customer and technical support experts are here to help! Please try again or contact Customer Service. Please contact Customer Service to unlock your account. Please try again or contact Customer Service. Please request another reset link. Our records indicate that this email address is already registered. We provide medical information and facilitate research collaborations. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. © 2021 Promega Corporation. Contains GoTaq® G2 enzyme. The pGEM®-5Zf(+) Vector serves as a standard cloning vector and as a template for in vitro transcription. You have not verified your email address. Note: You will not be able to access your account until your email is verified. Please update your browser to Internet Explorer 11 or above. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No.