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. Video Protocols. The pGEM®-T Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. X65308). 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. 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. Wysłaliśmy na podany adres e-mail do weryfikacji.
X65308). Wysokowydajna polimeraza DNA Taq do codziennych potrzeb PCR. Ratios from 3:1 to 1:3 provide good initial parameters. Login / Register Order Menu. Podaj nazwę użytkownika, aby otrzymać link do zresetowania hasła. Skontaktuj się z najbliższym przedstawicielem naukowym, Catalog number selected:
However, ratios of 8:1 to 1:8 have been used successfully. The pGEM®-T Easy pre-linearized Vector contains 3´-T overhangs at the insertion site to provide a compatible overhang for PCR products. Are there any tools that can assist with primer design for DNA sequencing? Regarding the pGEM-T vector I agree with Syed, you can insert the PCR fragment via T-A cloning. 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. Quick Protocols. Usage Suggestion：The ORF cDNA sequence can be amplified by PCR with M13-47 and RV-M primers. Wystąpił błąd w czasie zmiany hasła. 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 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. X65308). E-mail weryfikujący został wysłany na adres podany podczas rejestracji. Wystąpił błąd w czasie tworzenia konta. © 2007-2021 Sino Biological Inc. All rights reserved, Common Cytokine Receptor Signaling Pathway. 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. Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. The coding sequence was inserted by TA cloning. Video Protocols. Nie można otworzyć konto bez weryfikacji adresu e-mail. Please update your browser to Internet Explorer 11 or above. Proszę spróbować ponownie lub skontaktować się z Działem Obsługi Klienta. Proszę skontaktować się z Działem Obsługi Klienta, aby odblokować konto. Protocols. Quick Protocols. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Stay notified of Promega events, products and news. Alternatively, a double digestion may be used to release the insert from the vector. The provided 2X Rapid Ligation Buffer allows reactions to be completed in 1 hour at room temperature. 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. Both the pGEM®-T and pGEM -T Easy Vector contain multiple restriction sites within the multiple cloning region. 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. See Protocol for detailed storage recommendations. Twoje konto zostało utworzone. 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 position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). Polityka prywatności i przetwarzania danych
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. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Figure 1. Let's find the product that meets your needs. 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. When you select your country, you agree that we can place these functional cookies on your device. Protocols. 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. PROD | u7.5.14. pGEM-T easy plasmid DNA (500 ng, Promega, Madison, WI, USA) was then added and incubated for 1 h at 37 °C. Our website uses functional cookies that do not collect any personal information or track your browsing activity. + Datasheet. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. Complete Protocol. A resource designed for scientists just embarking on their career, focusing on fundamental technologies and techniques. 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). pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. SampleTextSampleText。:victory:pGEM-T_easy_vector_sequence质粒序列.docxpGEM-T_easy_vector质粒序列.txtLasteditedbysilicareon2012-10-18at17:39]
A3600. Thus, several options exist to remove the desired insert DNA with a single restriction digestion. Your professor will come around with the PGEM-T Easy Vector and T4 DNA ligase. TOP10, DH5α and TOP10F´, JM109. 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 3.0kb in size and contains the ampicillin resistance gene for selection. Complete Protocol. パフォーマンス. a. The pGEM ®-T and pGEM ®-T Easy Vector Systems are convenient systems for the cloning of PCR products. Protocolos en Vídeo. pGEM-T vector backbone. The pGEM-T vector is 3.0kb in size and contains the ampicillin resistance gene for selection. Determine the volume of PCR product to add to the ligation. Następnie należy skontaktować się z Działem Obsługi Klienta w celu odblokowania konta. Protocolos. The multiple cloning site is flanked by recognition sites for the restriction enzyme BstZI, allowing release of the insert by a single-enzyme digestion. We offer numerous convenient solutions to meet your lab's needs. Wystąpił błąd podczas utwrozenia konta. w10.0.13 | c1.0.0.2. 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 vector allows preparation of single-stranded DNA due to its f1 Origin of Replication. 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. 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 … If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Nie zweryfikowano podanego adresu e-mail. X65308). The pGEM®-T Vector is derived from the pGEM®-5Zf(+) Vector (GenBank® Accession No. Aby chronić Twoją prywatność, Twoje konto zostało zablokowane po 6 nieudanych próbach zalogowania się. The pGEM®-T Vector System II contains JM109 Competent Cells in addition to all of the pGEM®-T Vector System I components. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. Please try again or contact Customer Service. The concentration of PCR product Most commercially available competent cells are appropriate for the plasmid, e.g. Your commerce experience may be limited. Especificaciones. + Sequence information. The pGEM control and M13 primer provided in the kit should be used for troubleshooting purposes. The incubation period may be extended to increase the number of colonies after transformation. 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. Reactions using this buffer may be incubated for 1 hour at room temperature. .. Nicking of DNA was evaluated by ethidium bromide staining after electrophoresis separation in 0.8% agarose gels [ , ].
pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. This addition enables the ‘easy’ restriction of the plasmid for routine cloning applications, hence the name. Proszę sprawdzić połączenie internetowe i spróbować rejestracji ponownie. 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. Podany e-mail posiada już istniejące konto. Wystąpił błąd weryfikacji adresu e-mail. The coding sequence was inserted by TA cloning. If initial experiments with your PCR product are suboptimal, ratio optimization may be necessary. + Compare & Order pGEM-T vector backbone products + TOP customer support. The pGEM is a control template that can be used to isolate issues with sample quality, thermal cycler, kit or sequencing reaction purification. PCR cloning system for expression in mammalian cells. We provide medical information and facilitate research collaborations. Your password reset link has expired. E-mail z linkiem do zresetowania hasła został wysłany na adres podany podczas rejestracji. All Rights Reserved. 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。 Spróbuj ponownie lub skontaktuj się z Obsługą Klienta. Wysokowydajna polimeraza Taq z niezawierającymi Mg buforami reakcyjnymi. ベクターのT突出末端の安定性. EVOcards. Weryfikacja adresu e-mail jest niezbędna do utworzenia konta na promega.com. For clarity equivalent sequences in both constructs are only shown for pL4-GA Neo. X65308). Feature Options. Please request another reset link. Promega GmbH General Terms and Conditions of Business. Sprawdź swoją pocztę e-mail, aby potwierdzić adres e-mail. 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. ベクターマップ＆シークエンス. Wysokowydajna polimeraza DNA Taq w gotowej do użycia mieszaninie Master Mix. Protocolos Rápidos. 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 -. クイックプロトコル (pGEM-T Vectors) 製品マニュアル. © 2021 Promega Corporation. The pGEM®-T Easy Vector Systems are convenient systems to clone PCR products generated by certain thermostable polymerases. Specifications. pGEM®-T Vector Map and Sequence The pGEM®-T Vector is derived from the pGEM®-5Zf (+) Vector (GenBank® Accession No. The pGEM ®-T and pGEM ®-T Easy Vector Systems include a 2X Rapid Ligation Buffer for ligation of PCR products. Ratios from 3:1 to 1:3 provide good initial parameters. 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). I, pGEM-T Easy with a cloned genomic fragment comprising TcADK4 , ISs (solid bold lines) and flanking coding sequences (light grey boxes). pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual PDF (548 KB) – English. Trademarks
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. Legal and Trademarks
Complete Protocol. Proszę spróbować ponownie lub skontaktować się z Obsługą Klienta. 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. This product is available through the Promega Helix onsite stocking program. 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. These polymerases often add a single deoxyadenosine, in a template-independent fashion, to the 3´-ends of the amplified fragments. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt). However, ratios of 8:1 to 1:8 have been used successfully. There was an issue logging into your account. 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. We've detected that you are using an older version of Internet Explorer. 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." Gotowa do użycia zoptymalizowana mieszanina Master Mix do składania PCR w temperaturze pokojowej. Gratulacje! Read 3 answers by scientists to the question asked by Muh.Chaeril Ikramullah on Mar 3, 2021 Specifications. pGEM®-T and pGEM®-T Easy Vector Systems Technical Manual, pGEM T and pGEM T Easy Vector Systems FB033, 2017
The pGEM-T Easy vector has EcoRI restriction sites surrounding the proposed insert site, whereas the pGEM-T vector does not. The pGEM-T vector is a high-efficiency TA cloning vector which contains multiple cloning sites as shown below. 製品マニュアル（日本語） DH5α使用説明書. Dziękujemy za potwierdzenie adresu e-mail. pGEM T and pGEM T Easy Vector Systems FB033 PDF (202 KB) – English. Aby chronić Twoją prywatność, konto zostanie zablokowane po 6 nieudanych próbach. 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. The pGEM ® -T and pGEM ® -T Easy Vector Systems are convenient systems for the cloning of PCR products. XX CC pGEM-T has dT, which improves efficiency of ligation of PCR product. 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 Vector was created by linearizing the pGEM®-5Zf (+) Vector with EcoRV at base 51 and adding a T to both 3´-ends. The position of the T is indicated by * in the pGEM®-T Vector Sequence (.txt).