Akhileshwar Singh

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Questions related from Akhileshwar Singh

Why sometimes there is no expression of gateway vector cloned gene in plants?

When performing Gateway vector cloning and transforming Agrobacterium cells for plant transformation or transient expression of a gene of interest in tobacco leaves: Is it necessary to generate...

last month 3,128 0 View

What are the recommended strategies for purifying active recombinant proteins expressed in inclusion bodies for use in enzyme activity assays?

I am expressing several genes using the pET28(a) vector in E. coli BL21 cells. While some recombinant proteins are expressed, they accumulate predominantly in inclusion bodies. Although I purify...

04 May 2025 5,424 3 View

Why my cloned PCR products shows negative colonies?

I am trying to clone a few genes into the pENTR/D-TOPO vector. To do this, I use a previously cloned plasmid as a template for PCR product amplification. The PCR product is successfully amplified,...

23 April 2025 9,120 6 View