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Questions related from Hyunbae Lee
Hello. I am planning to utilize Chlamydomonas reinhardtii and GFP for my experiments. But they move so fast I cannot take pictures of them under microscope. Also, they seem to have...
22 February 2021 4,346 1 View
Hello. I am planning to work on with C. reinhardtii. I am going to use GFP vector to utilize fluorescent microscope. But one of researchers in our lab told me they moves very fast and it's hard...
18 February 2021 4,659 0 View
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 4,564 3 View
I am using plant protoplasts and polyethylene glycol (PEG) for transfection. Then I became curious if Ti-plasmid purified from Agrobacterium can be transfected directly by PEG mediation and can be...
03 February 2021 8,680 18 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 2,133 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 10,055 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 3,255 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 3,342 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 9,436 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 7,300 0 View
I am trying to look for a way to analyse profilin content in different tomato fruit tissues (peels, flesh and seeds). It seems the ideal choice is to run LS-MS/MS. Our lab is working on DNA, not...
05 June 2020 641 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 7,417 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 1,621 0 View
I wish to insert a very short DNA fragment into a vector. Since the DNA fragment doesn't have any proper stop codons, I want to add stop codons next to it. I could use the DNA fragment starting...
06 April 2020 8,989 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 9,714 1 View
I'm using petunia for protoplast extraction and gene transformation. It's been subcultured many many times and I guess the plants have been used for more than 3-4 years. We find it difficult to...
02 March 2020 3,153 2 View
I am growing protoplasts extracted from leaves in a liquid medium. They've formed colonies that consist of cells without cell walls. Now I need to count cell numbers... So I tried the same enzyme...
30 January 2020 879 3 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 3,917 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 819 3 View
