A cloning vector is a small piece of dna that can be stably maintained in an organism, and into which a foreign dna fragment can be inserted for cloning purposes. There is no genbank accession number for the pgem t easy vector. Insert transferred from pgemt easy vector to pgl3basic vector. The linearized ptg19 t vector with 3dt overhangs prevent vector recircularization, therefore resulting in high percentage of recombinant clones and low background. Wait a little while the tool is working and save the result. Quick protocol 1 pgem t and pgem t easy vector systems instructions for use of products a60, a80, a3600 and a3610. I tried to transform the ligated vector into li dh5 alpha by electroporation but got no results. The ptg19 t vector is designed for rapid and efficient cloning of pcr products with 3da overhangs. The pgemt easy vector systems are convenient systems for cloning pcr. They offer all of the advantages of the pgem t vector systems with the added convenience of recognition sites for ecori and noti flanking the insertion site. The pgem t vector systems are convenient systems to clone pcr products generated by certain thermostable polymerases.
Briefly centrifuge the pgem t or pgem t easy vector and control insert dna. Use only the t4 dna ligase provided with the system, as this has been tested for minimal exonuclease activity. The t overhangs at the insertion site greatly improve the efficiency of. Pcr cloning is a rapid method for cloning genes, and is often used for projects that require higher throughput than traditional cloning methods can accommodate. They offer all of the advantages of the pgem t vector systems with ecori and noti sites flanking the insertion site. Peter hegemanns lab contains the insert humanized photoactivated adenylyl cyclase and is published in j biol chem. Pgemt is a linearized cloning vector that can not be multiplied. Followed the ligation protocol as per promega instuctions. Receive the latest news, hot plasmids, discounts and more. Researchers have tried to develop pgem t like t vector. The pgem t and pgem t easy vector systems are convenient systems for the cloning of pcr products.
Have questions about your order, deposit, or a plasmid. Transfer the aqueous phase to the new phase lock tubes. Choose high quality vectors and click here for instant download. Instructions for use of products a60, a80, a3600, a3610. Convenient readytouse linearized 3dt overhang ptg19 t vector. Hipdf is a tool that will help users convert pdf to various file formats, vice versa. The technique relies on the ability of adenine a and thymine t complementary basepairs on different dna fragments to hybridize and, in the presence of ligase, become ligated together. This page is informational only this vector is not available from addgene. Protocol for ligations using the pgem t and pgem t easy vectors and the 2x rapid ligation buffer 3.
Promega has provide with a positive control insert along with the kit. These single 3 t overhangs at the insertion site greatly improve the efficiency of ligation of a pcr product into the plasmid. Welcome to vector database vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Pcr cloning differs from traditional cloning in that the dna fragment of interest, and even the vector, can be amplified by the polymerase chain reaction pcr and ligated together, without the use of restriction enzymes. Briefly centrifuge the pgem t or pgem t easy vector and control insert dna tubes to collect contents at the bottom of the tube. The pgemt easy vector systems are convenient systems to clone pcr products generated by certain thermostable polymerases. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. A versatile zero background tvector system for gene. The pgem t easy vector systems are convenient systems for cloning pcr products. Ta cloning also known as rapid cloning or t cloning is a subcloning technique that avoids the use of restriction enzymes and is easier and quicker than traditional subcloning. 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. Learn about the latest plasmid technologies and research tools.
Promega pgemt and pgemt easy vector systems pgem t easy. They claimed that their created new version pgem ft for t a cloning has a better performance then pgem t vector. We evaluated the cloning efficiency of different size pcr products into three t vector cloning systems. This is a free resource for the scientific community that is compiled by addgene.
The pgem t and pgem t easy vectorsa,b are linearized vectors with a single 3. How to multiply pgem t easy vector in the laboratory. The pgem t easy vector systems are convenient systems to clone pcr products. This is a free resource for the scientific community that is compiled by addgene this page is informational only this vector is not available from addgene please contact the manufacturer for further details. Get cells out of freezer, thaw on ice for 5 minutes 2. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide.
Comparing cloning efficiency of the pgemt and pgemt easy. Before being able to use the pgem t easy vector does it need to be digested so its linearized or is. The pgem t and pgem t easy vector systems ii include jm109 competent cells, high efficiency. The improved tvector system takes advantage of the restriction enzyme xcmi to. I have been trying to subclone an insert having 2300 bp into pgem t easy vector.
Riboprobe synthesis for in situ hybridization martindale lab. A versatile zero background tvector system for gene cloning and. The cloning vector may be dna taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. Download 264 latest free vector from freedesignfile. I need to know what are the differences between both pgemt and pgemt easy vector. These polymerases often add a single deoxyadenosine, in a templateindependent fashion, to the 3. Why do we always need cloning into pgem for example. The aim of molecular cloning is to insert the gene of interest into a plasmid vector, which is then inserted into a cell that will express the protein encoded by the gene of interest. To convert pdf to vector format, it is necessary to convert a pdf to bitmap image firstly and then you can easily convert the images to vectors. Mar072008 this sounds dumb and i think i know the answer but looking at the vector map has me confused. The pgemt easy vector systems are convenient systems for cloning pcr products.
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