Pgem vector pdf sample

Used as a standard cloning vector, a template for in vitro transcription or for production of. Here you can download a freebies sample pack, so you get a taste of their design stuff. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. Get cells out of freezer, thaw on ice for 5 minutes 2.

A versatile zero background tvector system for gene cloning and. Followed the ligation protocol as per promega instuctions. The pgem 9zf vector is a recombinant plasmid designed to provide a versatile range of cloning strategies, and contains t7 and sp6 rna polymerase promoters flanking a multiple cloning region within the. I tried to transform the ligated vector into li dh5 alpha by electroporation but got no results. T overhangs at the insertion site greatly improve the efficiency. Gel analysis and purification using ibis gelpcr dna fragment extraction kit 1. Construction of a high efficiency pcr products cloning t vector. Free vector samples by the talented design team of arsenalgomedia design studio. The highcopynumber pgemt and pgemt easy vectors contain t7 and. Genetic engineering is used in thousands of laboratories around the world. Insert transferred from pgemt easy vector to pgl3basic vector. The pgemt vector systems are convenient systems for cloning pcr products.

Learn about the latest plasmid technologies and research tools. A standard cloning vector used as a template for in vitro transcription or for production of circular ssdna. The primers i used have snabl and notl site in forward and reverse primer end respectively. In addition, the vector contains both the sp6 and t7 rna. Yes you can insert any gene sequence in pgemt easy vector but one thing you. The pgem t and pgem t easy vector systems are convenient systems for the cloning of pcr products. Promega pgemt and pgemt easy vector systems pgem t easy. The pgem 3z vector is intended for use as a standard cloning vector, as well as for the highly efficient synthesis of rna in vitro. A standard cloning vector that can be used for in vitro transcription and produce ssdna. If initial experiments with your pcr product are suboptimal, ratio optimization may be necessary. These polymerases often add a single deoxyadenosine, in a templateindependent fashion, to the 3. Have questions about your order, deposit, or a plasmid. May i know if it is true that gene with any sequence also can be inserted into pgemt easy.

The plasmid serves as a standard cloning vector, as a template for in vitro transcription and as a template for production of circular ssdna. Reactions using this buffer may be incubated for 1 hour at room temperature. The toverhangs at the insertion site greatly improve the efficiency of. Taq polymerase leaves an extra t or a at the end of the amplified dna in the pcr reaction, so the pcr product ligates readily. Since it has atoverhang, so will this affect the type of gene. We tested the promega t4 dna ligase, which is included in the pgemt easy. I have been trying to subclone an insert having 2300 bp into pgem t easy vector. Ligating the dna to yield a plasmid containing the geneofinterest. The pgemt vector systems are convenient ta vector systems for quickly cloning. Receive the latest news, hot plasmids, discounts and more. We evaluated the cloning efficiency of different size pcr products into three tvector cloning systems.

Promega has provide with a positive control insert along with the kit. How to multiply pgem t easy vector in the laboratory. The mcs of the pgemt easy vector contains sequences on either side of the insert that are recognized by the restriction enzymes not i and ecor i. The pgem t and pgem t easy vectorsa,b are linearized vectors with a single 3. I find that it is more efficient than pgemt easy and that i get more white colonies with my insert. The only difference between pgemt and pgemt easy is in the multiple cloning site mcs. The incubation period may be extended to increase the number of colonies after transformation. Why do we always need cloning into pgem for example. In addition, the vector contains both the sp6 and t7 rna polymerase promoters flanking the. Why do we always need cloning into pgem for example vector, before. This vector contains t7 and sp6 rna polymerase promoters flanking a multiple cloning region within the.

They claimed that their created new version pgemft for ta cloning has a better performance. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. These single 3t overhangs at the insertion site greatly improve the efficiency of ligation of a pcr product into the plasmid. This allows the insert dna to be removed with a single restriction digest using either of these enzymes. Researchers have tried to develop pgemtlike tvector.