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®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Your password reset link has expired. Wystąpił błąd weryfikacji adresu e-mail. + Compare & Order pGEM-T vector backbone products + TOP customer support. X65308). 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). Quick Protocols. 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 is a control template that can be used to isolate issues with sample quality, thermal cycler, kit or sequencing reaction purification. 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. However, ratios of 8:1 to 1:8 have been used successfully. The provided 2X Rapid Ligation Buffer allows reactions to be completed in 1 hour at room temperature. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Your commerce experience may be limited. + Datasheet. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. 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. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Wystąpił błąd w czasie zmiany hasła. Primer3 is a great tool to pick your primers from a particular sequence. The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. Wysokowydajna polimeraza Taq z niezawierającymi Mg buforami reakcyjnymi. The pGEM®-T Easy pre-linearized Vector contains 3´-T overhangs at the insertion site to provide a compatible overhang for PCR products. Twoje konto zostało utworzone. The concentration of PCR product However, ratios of 8:1 to 1:8 have been used successfully. Figure 1. Usage Suggestion：The ORF cDNA sequence can be amplified by PCR with M13-47 and RV-M primers. We've detected that you are using an older version of Internet Explorer. For clarity equivalent sequences in both constructs are only shown for pL4-GA Neo. Please try again or contact Customer Service. Login / Register Order Menu. Stay notified of Promega events, products and news. Nie można otworzyć konto bez weryfikacji adresu e-mail. This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. 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. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. The vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. Ratios from 3:1 to 1:3 provide good initial parameters. Protocolos. Proszę sprawdzić połączenie internetowe i spróbować rejestracji ponownie. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). © 2007-2021 Sino Biological Inc. All rights reserved, Common Cytokine Receptor Signaling Pathway. Wystąpił błąd podczas utwrozenia konta. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. 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. Sprawdź swoją pocztę e-mail, aby potwierdzić adres e-mail. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Protocolos en Vídeo. Następnie należy skontaktować się z Działem Obsługi Klienta w celu odblokowania konta. + Sequence information. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. Both the pGEM®-T and pGEM -T Easy Vector contain multiple restriction sites within the multiple cloning region. Video Protocols. a. E-mail z linkiem do zresetowania hasła został wysłany na adres podany podczas rejestracji. 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. Please request another reset link. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. Proszę spróbować ponownie lub skontaktować się z Działem Obsługi Klienta. This product is available through the Promega Helix onsite stocking program. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. 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 … Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. However, ratios of 8:1 to 1:8 have been used successfully. Insertional inactivation of the α-peptide allows recombinant clones to be directly identified by Blue/White Screening on indicator plates. 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. ベクターマップ＆シークエンス. Quick Protocols. The incubation period may be extended to increase the number of colonies after transformation. Please update your browser to Internet Explorer 11 or above. 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. PCR cloning system for expression in mammalian cells. Determine the volume of PCR product to add to the ligation. X65308). 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 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。 パフォーマンス. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. EVOcards. Protocols. Podaj nazwę użytkownika, aby otrzymać link do zresetowania hasła. 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. Aby chronić Twoją prywatność, Twoje konto zostało zablokowane po 6 nieudanych próbach zalogowania się. Legal and Trademarks XX CC pGEM-T has dT, which improves efficiency of ligation of PCR product. Ratios from 3:1 to 1:3 provide good initial parameters. We offer numerous convenient solutions to meet your lab's needs. 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. X65308). 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. All Rights Reserved. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). The pGEM ®-T and pGEM ®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. Especificaciones. 製品マニュアル（日本語） DH5α使用説明書. 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. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. Our website uses functional cookies that do not collect any personal information or track your browsing activity. A3600. See Protocol for detailed storage recommendations. We provide medical information and facilitate research collaborations. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Feature Options. Complete Protocol. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Weryfikacja adresu e-mail jest niezbędna do utworzenia konta na promega.com. ベクターのT突出末端の安定性. Wysokowydajna polimeraza DNA Taq w gotowej do użycia mieszaninie Master Mix. Trademarks Podany e-mail posiada już istniejące konto. 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. Procedure: 1. Are there any tools that can assist with primer design for DNA sequencing? Promega GmbH General Terms and Conditions of Business. X65308). Video Protocols. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. Benefit from the greatest possible flexibility in the choice of handling and managing your sequencing primers. 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. Read 3 answers by scientists to the question asked by Muh.Chaeril Ikramullah on Mar 3, 2021 PCR cloning vectors with 3 options for insert excision. E-mail weryfikujący został wysłany na adres podany podczas rejestracji. Specifications. I, pGEM-T Easy with a cloned genomic fragment comprising TcADK4 , ISs (solid bold lines) and flanking coding sequences (light grey boxes). X65308). Gratulacje! There was an issue logging into your account. Complete Protocol. pGEM-T Vector Information Description The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. pGEM-T vector backbone. Let's find the product that meets your needs. When you select your country, you agree that we can place these functional cookies on your device. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Your professor will come around with the PGEM-T Easy Vector and T4 DNA ligase. TOP10, DH5α and TOP10F´, JM109. Reactions using this buffer may be incubated for 1 hour at room temperature. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Most commercially available competent cells are appropriate for the plasmid, e.g. Wysokowydajna polimeraza DNA Taq do codziennych potrzeb PCR. 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 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 incubation period may be extended to increase the number of colonies after transformation. Regarding the pGEM-T vector I agree with Syed, you can insert the PCR fragment via T-A cloning. Wysłaliśmy na podany adres e-mail do weryfikacji. w10.0.13 | c1.0.0.2. Nie zweryfikowano podanego adresu e-mail. .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ]. クイックプロトコル (pGEM-T Vectors) 製品マニュアル. 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. 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. Polityka prywatności i przetwarzania danych 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 -. © 2021 Promega Corporation. Wystąpił błąd w czasie tworzenia konta. 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 has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual, pGEM T and pGEM T Easy Vector Systems FB033, 2017 Complete Protocol. Gotowa do użycia zoptymalizowana mieszanina Master Mix do składania PCR w temperaturze pokojowej. Specifications. 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. Proszę skontaktować się z Działem Obsługi Klienta, aby odblokować konto. The coding sequence was inserted by TA cloning. 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. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. Aby chronić Twoją prywatność, konto zostanie zablokowane po 6 nieudanych próbach. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Protocols. The pGEM ® -T and pGEM ® -T Easy Vector Systems are convenient systems for the cloning of PCR products. The pGEM control and M13 primer provided in the kit should be used for troubleshooting purposes. PROD | u7.5.14. The coding sequence was inserted by TA cloning. Proszę spróbować ponownie lub skontaktować się z Obsługą Klienta. Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. Alternatively, a double digestion may be used to release the insert from the vector. 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. REQUIRED MATERIALS PGEM-T Easy plasmid (Kit ordered from Fisher PR-1380) 2x rapid ligation buffer T4 DNA Ligase enzyme **Note a few ingredients to the Ligation reaction are NOT on your desk due to the very small volumes needed. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. https://www.snapgene.com/.../?set=basic_cloning_vectors&plasmid=pGEM-T Dziękujemy za potwierdzenie adresu e-mail. SampleTextSampleText。:victory:pGEM-T_easy_vector_sequence质粒序列.docxpGEM-T_easy_vector质粒序列.txtLasteditedbysilicareon2012-10-18at17:39] If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary.
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