The first step in Gateway cloning is the preparation of a Gateway Entry clone. There are a few different ways to make entry clone. Gateway attB1 and attB2 sequences are added to the 5' …

Gateway cloning is a highly efficient alternative to restriction cloning and does not require the use of restriction enzymes. Instead, an insert is moved into a vector through a two-step …

The attB1 site is indicated in bold and the ATG initiation codon for the protein of interest is underlined. Inclusion of the Shine-Dalgarno and Kozak consensus sequence allows protein …

Recombination of attP and attB sites creates attL and attR sites. Gateway vectors contain modified versions of the att sites so that scientists can easily clone in their desired DNA …

您可以利用这些载体克隆含有attB序列 (InvitrogenTM pDONRTM 载体) 或特异限制性位点 (InvitrogenTM pENTRTM 载体) 的引物扩增的PCR 产物。 使用PCR 生成入门克隆,两个短的人 …

nucleotide changes within the 25-bp recognition region (Fig. 1C) has been used to engineer different subtypes of att sites that recombine only with each other (i.e., attB1 sites with attP1 …

What are the sequences of the attB1 and attB2 sites that I need to add to my PCR product for Gateway® cloning?

Learn how to simulate Gateway® cloning in Geneious Prime, including how to automatically design attB-linked oligonucleotide primers, simulate BP and LR reactions. Geneious Prime …

Jan 11, 2021 · After second-strand synthesis and end polishing reactions a phosphorylated attB1 adapter is ligated to the 5′ end of the cDNA. The presence of biotin prevents the attB1 adapter …

Set whether the insert is recombined in the forward (attB1 → Insert → attB2) or reverse (attB2 → Insert → attB1) orientation. Set the desired target melting temperature (T m) for the new PCR …