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Questions related from Hyunbae Lee
Hello, I am planning to insert GOI between CaMV 35 promoter and mGFP in pCambia1302 to produce a fusion protein and see subcellular localization of the protein. But the vector says it's membrane...
16 February 2021 4,529 3 View
Hello, I was planning to modify 4 kb GFP plasmid for transient expression in plants. I was going to add 4 kb gene and make 8 kb plasmid. Our lab use plant protoplasts for experiments and when I...
06 November 2020 2,121 3 View
Hello, I am interested in producing purified Cas9 in my lab. But we are not dealing with proteins so we don't have proper settings for protein purification. It seems a step using chromatography is...
26 October 2020 10,042 3 View
I isolated and purified plasmid DNA from my E. coli culture using protocol of Addgene. They say overnight culture at -20 °C would help precipitation of plasmid DNA in case of using 70% alcohol for...
23 October 2020 3,238 3 View
Hello. I am planning an RNAi experiment. I want to silence my gene of interest using RNAi. My gene of interest is crucial for plant development. Also, my gene of interest is known to function even...
22 September 2020 3,327 3 View
I used up GFP plasmid in our lab so I am trying to transform GFP plasmid into E. coli then extract GFP plasmid after culture. I did transformation with this protocol 1. thawing E. coli in ice...
25 August 2020 9,429 2 View
We use protoplasts for experiment and I found some things are moving inside the cells. What are these things??? Are chloroplasts and mitochondria mobile??? Or can it be some kind of bacteria? I...
23 July 2020 7,293 0 View
I'm trying to transform GFP plasmids to E. coli and clone them. We tried it but the concentration is too low, like 100ug/ul. We use 50ug of GFP plasmids for each experiment and the concentration...
12 May 2020 7,408 9 View
I use PEG for gene transformation of petunia protoplasts. I use PEG4000... When washing is done, the cells aggregate and eventually die. I guess it's because the concentration of the cells are...
16 April 2020 1,614 0 View
I'm doing GFP transformation using petunia protoplasts. I've tried 6 times and it seems I succeeded it for the first time. I guess it's important to keep cells alive so I changed the protocol a...
02 March 2020 9,703 1 View
I am a graduate student majoring in horticulture. My first semester just ended and I'm planning to design a simple experiment. I want to make synthetic DNA that would encodes a short peptide that...
27 December 2019 3,895 1 View
I am going to transfect plant protoplasts with pCambia1302 for stable expression of GOI. I am not used to the use of plasmid and don't really understand how a single vector can introduce a gene to...
01 January 1970 808 3 View
