The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. EVOcards. The coding sequence was inserted by TA cloning. Your password reset link has expired. パフォーマンス. Protocols. X65308). I, pGEM-T Easy with a cloned genomic fragment comprising TcADK4 , ISs (solid bold lines) and flanking coding sequences (light grey boxes). Protocolos. PCR cloning vectors with 3 options for insert excision. Trademarks 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. pGEM-T vector backbone. Wysokowydajna polimeraza DNA Taq do codziennych potrzeb PCR. Video Protocols. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). PCR cloning system for expression in mammalian cells. Aby chronić Twoją prywatność, Twoje konto zostało zablokowane po 6 nieudanych próbach zalogowania się. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual, pGEM T and pGEM T Easy Vector Systems FB033, 2017 Feature Options. Proszę spróbować ponownie lub skontaktować się z Obsługą Klienta. XX CC pGEM-T has dT, which improves efficiency of ligation of PCR product. X65308). Your professor will come around with the PGEM-T Easy Vector and T4 DNA ligase. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. 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. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. Ratios from 3:1 to 1:3 provide good initial parameters. Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. w10.0.13 | c1.0.0.2. We provide medical information and facilitate research collaborations. Usage Suggestion：The ORF cDNA sequence can be amplified by PCR with M13-47 and RV-M primers. Complete Protocol. Please try again or contact Customer Service. Proszę spróbować ponownie lub skontaktować się z Działem Obsługi Klienta. This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. Specifications. Are there any tools that can assist with primer design for DNA sequencing? CC NM (pGEM-T) CC CM (yes) CC NA (ds-DNA) CC TP (circular) CC ST () CC TY (phagemid) CC SP (Promega) CC HO (E.coli) CC CP () CC FN (cloning)(transcription) CC SE (color blue/white) CC PA (pGEM-5Zf+) CC BR () CC OF () CC OR () XX FH Key Location/Qualifiers FH FT misc_feature 0..0 FT /note="1. pGEM-5Zf+ 3003bp FT -> pGEM-T … 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. The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. Login / Register Order Menu. The coding sequence was inserted by TA cloning. The pGEM is a control template that can be used to isolate issues with sample quality, thermal cycler, kit or sequencing reaction purification. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. The pGEM ®-T and pGEM ®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. Proszę sprawdzić połączenie internetowe i spróbować rejestracji ponownie. When you select your country, you agree that we can place these functional cookies on your device. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Wystąpił błąd weryfikacji adresu e-mail. Wystąpił błąd podczas utwrozenia konta. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. SampleTextSampleText。:victory:pGEM-T_easy_vector_sequence质粒序列.docxpGEM-T_easy_vector质粒序列.txtLasteditedbysilicareon2012-10-18at17:39] The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Benefit from the greatest possible flexibility in the choice of handling and managing your sequencing primers. 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. Determine the volume of PCR product to add to the ligation. The pGEM®-T Easy pre-linearized Vector contains 3´-T overhangs at the insertion site to provide a compatible overhang for PCR products. Complete Protocol. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). 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. We've detected that you are using an older version of Internet Explorer. Promega GmbH General Terms and Conditions of Business. Primer3 is a great tool to pick your primers from a particular sequence. 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. Let's find the product that meets your needs. See Protocol for detailed storage recommendations. The incubation period may be extended to increase the number of colonies after transformation. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. Wysłaliśmy na podany adres e-mail do weryfikacji. Twoje konto zostało utworzone. Weryfikacja adresu e-mail jest niezbędna do utworzenia konta na promega.com. The incubation period may be extended to increase the number of colonies after transformation. Legal and Trademarks Wysokowydajna polimeraza DNA Taq w gotowej do użycia mieszaninie Master Mix. Polityka prywatności i przetwarzania danych Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. © 2007-2021 Sino Biological Inc. All rights reserved, Common Cytokine Receptor Signaling Pathway. Please update your browser to Internet Explorer 11 or above. Your commerce experience may be limited. Especificaciones. クイックプロトコル (pGEM-T Vectors) 製品マニュアル. Protocolos Rápidos. Podaj nazwę użytkownika, aby otrzymać link do zresetowania hasła. E-mail weryfikujący został wysłany na adres podany podczas rejestracji. 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. Podany e-mail posiada już istniejące konto. 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. Gratulacje! Procedure: 1. 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. Dziękujemy za potwierdzenie adresu e-mail. X65308). 製品マニュアル（日本語） DH5α使用説明書. © 2021 Promega Corporation. 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. Wystąpił błąd w czasie zmiany hasła. What do you mean by " if you are going for expression from that gene then try to avoid pGEM-T easy vector because later these overhang can cause problem in expression level." a. However, ratios of 8:1 to 1:8 have been used successfully. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. Skontaktuj się z najbliższym przedstawicielem naukowym, Catalog number selected: 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. Nie można otworzyć konto bez weryfikacji adresu e-mail. pGEM-T Easy Vector: 3016 bp 1 1000 2000 3000 3016 ApaI (14) AatII (20) NcoI (37) SacII (49) SpeI (65) PstI (89) SalI (91) NdeI (98) SacI (110) M13_reverse_primer Sp6_primer M13_pUC_rev_primer lac_promoter ORF frame 3 Ampicillin AmpR_promoter f1_origin lacZ_a M13_pUC_fwd_primer M13_forward20_primer. Specifications. TOP10, DH5α and TOP10F´, JM109. 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. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Protocols. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. There was an issue logging into your account. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. Stay notified of Promega events, products and news. pGEM-T Vector Information Description The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). A3600. 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. Regarding the pGEM-T vector I agree with Syed, you can insert the PCR fragment via T-A cloning. Aby chronić Twoją prywatność, konto zostanie zablokowane po 6 nieudanych próbach. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. The provided 2X Rapid Ligation Buffer allows reactions to be completed in 1 hour at room temperature. 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. + Compare & Order pGEM-T vector backbone products + TOP customer support. However, ratios of 8:1 to 1:8 have been used successfully. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. 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. Nie zweryfikowano podanego adresu e-mail. https://www.snapgene.com/.../?set=basic_cloning_vectors&plasmid=pGEM-T However, ratios of 8:1 to 1:8 have been used successfully. II, TGRVs produced by replacement of a fragment of TcADK4 with the SMs of p Tc R-HG Hyg - and p Tc R-GA Neo -. Our website uses functional cookies that do not collect any personal information or track your browsing activity. 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. X65308). Video Protocols. Ratios from 3:1 to 1:3 provide good initial parameters. X65308). pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. 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.

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