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

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