The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Nie zweryfikowano podanego adresu e-mail. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. Legal and Trademarks Sprawdź swoją pocztę e-mail, aby potwierdzić adres e-mail. 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. Wystąpił błąd w czasie tworzenia konta. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. Procedure: 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. The incubation period may be extended to increase the number of colonies after transformation. Complete Protocol. Wysłaliśmy na podany adres e-mail do weryfikacji. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. a. The pGEM®-T Easy Vector multiple cloning region is flanked by recognition sites for the restriction enzymes EcoRI, BstZI and NotI, thus providing three single-enzyme digestions for release of the insert. For clarity equivalent sequences in both constructs are only shown for pL4-GA Neo. w10.0.13 | c1.0.0.2. Aby chronić Twoją prywatność, konto zostanie zablokowane po 6 nieudanych próbach. Please request another reset link. The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. 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 pGEM®-T Vector was created by linearizing the pGEM®-5Zf (+) Vector with EcoRV at base 51 and adding a T to both 3´-ends. Protocolos Rápidos. Wysokowydajna polimeraza Taq z niezawierającymi Mg buforami reakcyjnymi. https://www.snapgene.com/.../?set=basic_cloning_vectors&plasmid=pGEM-T Proszę sprawdzić połączenie internetowe i spróbować rejestracji ponownie. Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. Are there any tools that can assist with primer design for DNA sequencing? EVOcards. © 2007-2021 Sino Biological Inc. All rights reserved, Common Cytokine Receptor Signaling Pathway. Weryfikacja adresu e-mail jest niezbędna do utworzenia konta na promega.com. E-mail weryfikujący został wysłany na adres podany podczas rejestracji. Reactions using this buffer may be incubated for 1 hour at room temperature. I, pGEM-T Easy with a cloned genomic fragment comprising TcADK4 , ISs (solid bold lines) and flanking coding sequences (light grey boxes). Gratulacje! The pGEM ® -T and pGEM ® -T Easy Vector Systems are convenient systems for the cloning of PCR products. Proszę skontaktować się z Działem Obsługi Klienta, aby odblokować konto. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. Our website uses functional cookies that do not collect any personal information or track your browsing activity. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Most commercially available competent cells are appropriate for the plasmid, e.g. Podany e-mail posiada już istniejące konto. TOP10, DH5α and TOP10F´, JM109. The incubation period may be extended to increase the number of colonies after transformation. Aby chronić Twoją prywatność, Twoje konto zostało zablokowane po 6 nieudanych próbach zalogowania się. E-mail z linkiem do zresetowania hasła został wysłany na adres podany podczas rejestracji. Login / Register Order Menu. 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." The pGEM ®-T and pGEM ®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. There was an issue logging into your account. The promoter and multiple cloning sequence of the pGEM®-T (Panel A) and pGEM®-T Easy (Panel B) Vectors.The top strand of the sequence shown corresponds to the RNA synthesized by T7 RNA Polymerase.The bottom strand corresponds to the RNA synthe-sized by SP6 RNA Polymerase. Podaj nazwę użytkownika, aby otrzymać link do zresetowania hasła. X65308). クイックプロトコル (pGEM-T Vectors) 製品マニュアル. + Sequence information. 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。 The concentration of PCR product pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual, pGEM T and pGEM T Easy Vector Systems FB033, 2017 Complete Protocol. Protocols. Especificaciones. Protocolos en Vídeo. Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. Quick Protocols. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. Nie można otworzyć konto bez weryfikacji adresu e-mail. Primer3 is a great tool to pick your primers from a particular sequence. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). 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. 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. PCR cloning vectors with 3 options for insert excision. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. パフォーマンス. The pGEM is a control template that can be used to isolate issues with sample quality, thermal cycler, kit or sequencing reaction purification. pGEM-T vector backbone. pGEM®-T Vector Map and Sequence 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. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. © 2021 Promega Corporation. The coding sequence was inserted by TA cloning. Figure 1. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. Determine the volume of PCR product to add to the ligation. The provided 2X Rapid Ligation Buffer allows reactions to be completed in 1 hour at room temperature. ベクターマップ＆シークエンス. See Protocol for detailed storage recommendations. Wystąpił błąd weryfikacji adresu e-mail. However, ratios of 8:1 to 1:8 have been used successfully. Complete Protocol. X65308). We offer numerous convenient solutions to meet your lab's needs. Wystąpił błąd podczas utwrozenia konta. Protocols. Both the pGEM®-T and pGEM -T Easy Vector contain multiple restriction sites within the multiple cloning region. Feature Options. 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 pGEM®-T Vector was created by linearizing the pGEM®-5Zf (+) Vector with EcoRV at base 51 and adding a T to both 3´-ends. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Video Protocols. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). 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 pGEM®-T Easy pre-linearized Vector contains 3´-T overhangs at the insertion site to provide a compatible overhang for PCR products. Specifications. 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. Następnie należy skontaktować się z Działem Obsługi Klienta w celu odblokowania konta. The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. pGEM-T Vector Information Description The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. Quick Protocols. Gotowa do użycia zoptymalizowana mieszanina Master Mix do składania PCR w temperaturze pokojowej. Protocolos. Promega GmbH General Terms and Conditions of Business. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. 製品マニュアル（日本語） DH5α使用説明書. The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. 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. SampleTextSampleText。:victory:pGEM-T_easy_vector_sequence质粒序列.docxpGEM-T_easy_vector质粒序列.txtLasteditedbysilicareon2012-10-18at17:39] However, ratios of 8:1 to 1:8 have been used successfully. Ratios from 3:1 to 1:3 provide good initial parameters. Proszę spróbować ponownie lub skontaktować się z Obsługą Klienta. Twoje konto zostało utworzone. However, ratios of 8:1 to 1:8 have been used successfully. 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. Wystąpił błąd w czasie zmiany hasła. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. PROD | u7.5.14. .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. PLos ONE, Badania serologiczne SARS-CoV-2 i testy PCR, Badania w kierunku wirusów i rozwój szczepionek, 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, Polityka prywatności i przetwarzania danych, Promega GmbH General Terms and Conditions of Business, Insert excision with a BstZI single digest, Ligation can be completed in 1 hour at room temperature, Available with or without competent cells. Skontaktuj się z najbliższym przedstawicielem naukowym, Catalog number selected: X65308). + Datasheet. The pGEM control and M13 primer provided in the kit should be used for troubleshooting purposes. Alternatively, a double digestion may be used to release the insert from the vector. PCR cloning system for expression in mammalian cells. Polityka prywatności i przetwarzania danych XX CC pGEM-T has dT, which improves efficiency of ligation of PCR product. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. Wysokowydajna polimeraza DNA Taq do codziennych potrzeb PCR. Let's find the product that meets your needs. 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. Specifications. Regarding the pGEM-T vector I agree with Syed, you can insert the PCR fragment via T-A cloning. Your password reset link has expired. Wysokowydajna polimeraza DNA Taq w gotowej do użycia mieszaninie Master Mix. The pGEM-T Easy vector has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. Ratios from 3:1 to 1:3 provide good initial parameters. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Your professor will come around with the PGEM-T Easy Vector and T4 DNA ligase. X65308). + Compare & Order pGEM-T vector backbone products + TOP customer support. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). X65308). Trademarks Video Protocols. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Benefit from the greatest possible flexibility in the choice of handling and managing your sequencing primers. Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. Stay notified of Promega events, products and news. We've detected that you are using an older version of Internet Explorer. pGEM®-T Easy, pGEM-T Easy: Analyze: Sequence: Plasmid Type: Bacterial Expression: Expression Level: High: Cloning Method: Unknown: Size: 3015: 5' Sequencing 1 Primer: T7, SP6, M13Fwd or M13Rev: Bacterial Resistance: Ampicillin: Notes: The only difference between pGEM-T and pGEM-T Easy is in the multiple cloning site (MCS). When you select your country, you agree that we can place these functional cookies on your device.
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