Thank you for verifying your email address. A verification email has been sent to the primary email address associated with your account. The gold arrows indicate 5′-RACE products characterized from the Δ5G construct. Please try again or contact Customer Service. You've created a Promega.com account. Ligation Using 2X Rapid Ligation Buffer 1. A3600. Enter your username and we'll send a link to reset your password. Second-generation, high-performance GoTaq® G2 DNA Polymerase with Mg-free buffers. 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. Promega Notes 71 , 8–9. We offer numerous convenient solutions to meet your lab's needs. Background: Development of resistance to doxorubicin-based chemotherapy limits its curative effect in osteosarcoma. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. As a result, PCR products amplified using GoTaq® DNA Polymerase will contain A-overhangs which makes it suitable for T-vector cloning. Legal and Trademarks Abstract. 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 4/17 www.promega.com 2. There was an issue sending the verification email. We provide medical information and facilitate research collaborations. Trademarks. Instructions for Use of Product(s) A1360, A1380, A3600, A3610. However, ratios of 8:1 to 1:8 have been used successfully. Please try again or contact Customer Service. Alternatively, a double digestion may be used to release the insert from the vector. Please try again or contact Customer Service. The shoot apex tissues of young seedlings were fixed in RNase-free formalin/acetic acid/alcohol fixative. Please try again or contact Customer Service. 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. Please try again or contact Customer Service. Contains GoTaq® G2 enzyme. 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. (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. Literature # TM042. Please contact Customer Service to unlock your account. The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Get in touch with a nearby distributor or sales representative. PCR cloning system for expression in mammalian cells. PCR products with low concentration or generating heterogeneity in the sequencing chromatograms were cloned into pGEM-T Easy Vector (Promega) for sequencing. Reactions using this buffer may be incubated for 1 hour at room temperature. 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. 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 is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. The pGEM®-T Vector cloning region is flanked by recognition sites for the enzyme BstZI. 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. Stay notified of Promega events, products and news. A verified email address is required to access the full functionality of your Promega.com account. US orders: Ship Saturday March 13 for arrival on Monday March 15. Dismiss. Quick PROTOCOL 1 pGEM®-T and pGEM®-T Easy Vector Systems INSTRUCTIONS FOR USE OF PRODUCTS A1360, A1380, A3600 AND A3610. Due to our annual Physical inventory orders received after 5PM Central Time on Tuesday March 9 will be shipped according to the following schedule: We evaluated the cloning efficiency of different size PCR products into three T-vector cloning systems. 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. You have successfully reset your password. Second-generation, high-performance GoTaq® G2 DNA Polymerase in a ready-to-use master mix. 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. 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. You have successfully reset your password. There was an issue with the password reset process. A short summary of this paper. A password reset email has been sent to the primary email address associated with your account. Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. X65308). After that, you will need to contact Customer Service to unlock your account. Diese Seite wurde zuletzt am … The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. 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). 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 Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. This paper. The insertion site is flanked by BstZI sites. The pGEM®-5Zf(+) Vector serves as a standard cloning vector and as a template for in vitro transcription. A verification email has been sent to the primary email address associated with your account. Page 4 Revised 5/07 GGAGA GCTCC CAACG CGTTG GATGC ATAGC TTGAG … 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. See Protocol for detailed storage recommendations. In this study, we describe a method for producing armored L-RNA. Please try again or contact Customer Service. Your commerce experience may be limited. 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. 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. 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. Please contact Customer Service to unlock your account. Terms and Conditions 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 in vivo ECM is comprised not only of proteins but also of a variety of non-protein components. Molecular Cloning: A Laboratory Manual Third Edition. Revised 4/17 www.promega.com 2. Revised 12/18 www.promega.com 3. However, ratios of 8:1 to 1:8 have been used successfully. To protect your privacy, your account has been locked after 6 failed login attempts. Download Full PDF Package. PLos ONE, Plate Readers, Fluorometers & Luminometers, Save 20% on pGL4 Luciferase Reporter Vectors, enter PGL20 at checkout. Summary of Changes Introduction. Download PDF. The 3.9-kb product was cloned into pGEM-T Easy (Promega… The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). 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 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. Accordingly, as a means of enhancing tissue invasion, tumor cells use matrix metalloproteinases to degrade ECM proteins. We provide medical information and facilitate research collaborations. pGEM®-T Easy Parental vector for TA cloning of PCR products. Check your inbox to complete email verification. Our customer and technical support experts are here to help! A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. 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 logging into your account. Please try again or contact Customer Service. This is a free resource for the scientific community that is compiled by Addgene.. PDF (548k). 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. There was an issue resetting your password. www.promega.com Part# TM042 Printed in USA. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. There was an issue verifying your email address. Frackman, S. and Kephart, D (1999) Rapid ligation for the pGEM®-T and pGEM®-T Easy Vector Systems. This vector contains T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide coding region of β-galactosidase. Our records indicate that this email address is already registered. Please update your browser to Internet Explorer 11 or above. Congratulations! Canada orders: Ship Monday March 15 for arrival on Tuesday March 16. Enter your username and we'll send a link to reset your password. pGEM®-T Parental vector for TA cloning of PCR products. 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. This page is informational only - this vector is NOT available from Addgene - please contact the manufacturer for further details. The vector carries the lacZ alpha-peptide and the multiple cloning region arrangement from pUC18 allowing selection of recombinants by blue/white screening. There was an error processing your request. 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. Thank you for verifying your email address. There was an issue creating your account. The single major product was cloned into the pGEM-T-easy vector (Promega) and was sequenced. Ratios from 3:1 to 1:3 provide good initial parameters. ®Protocol for Ligations Using the pGEM -T and pGEM®-T Easy Vectors and the 2X Rapid Ligation Buffer 3.A. There was an issue with the password reset process. 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 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. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. ®Briefly centrifuge the pGEM-T or pGEM®-T Easy Vector and Control Insert … PCR cloning vectors with 3 options for insert excision. © 2021 Promega Corporation. 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. 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. Issai Falcon. You've created a Promega.com account. 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. Description. Welcome to Vector Database!. Privacy Policy and Requests for Information Our website uses functional cookies that do not collect any personal information or track your browsing activity. US orders: Ship Saturday March 13 for arrival on Monday March 15. RNase-resistant, noninfectious virus-like particles containing exogenous RNA sequences (armored RNA) are good candidates as RNA controls and standards in RNA virus detection. 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.
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