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Questions related from Yang Xiao
I did chromatin extraction using both commercial kit and homemade recipe. By using RNase A and protease K digestion together with phenol-chloroform-isoamyl or on-column cleaning, I then isolated...
23 July 2019 720 2 View
I used bisulfite conversion-->region-specific PCR-->cloning method to determine methylation of a gene at a specific region between two treatment groups. I had 8 individuals in one treatment...
21 January 2019 9,085 10 View
I extracted gDNA, amplified the target regions (554bp, 226bp, 344bp, and 194bp), gel extracted the target bands and purified them. Then I did the ligation (3:1 insert to vector) at 4 C overnight,...
13 August 2018 2,556 2 View
I cloned the PCR product (gel checked) into the p-GEM T Easy vector system and extracted plasmid DNA. Now I am doing restriction enzyme digestion to check if the target got successfully inserted....
10 July 2018 2,680 9 View
I recently isolated RNA from chicken abdominal adipocytes (induced from preadipocytes passaged 5 times). I previously only use cells passaged within 3 times for experiments, and the RNA yield and...
01 November 2017 1,570 2 View
I am trying to isolate the mature adipocytes directly from chicken abdominal adipose tissue, ceiling culture the adipocytes, and then isolate the RNA from these proliferated adipocytes. I followed...
29 August 2017 5,304 4 View
I am currently trying to isolate adipocytes from adipose tissue in juvenile chickens and want to use those adipocytes for RNA isolation. I found several articles and protocols online but they are...
21 August 2017 4,154 2 View
I am confused by the role of cAMP on adipogenesis and lipolysis. I know that when cAMP is high, it activates PKA and thus phosphorylates HSL and perilipin A to facilitate lipolysis, and when cAMP,...
28 April 2017 6,101 1 View
I am a bit confused about the primer design. If the same gene is expressed in different tissues, why should the primers be designed in a tissue-specific way instead of using the same primers to...
23 April 2017 6,713 7 View
I transduced the SVF preadipocytes with lentivirus and about a month after transduction and selection, now the survived cells are like this. For one like this I observed small lipid droplets at...
05 April 2017 7,443 2 View
We bought customized pLenti-shRNA-GFP with anti-blasticidin selection gene. I tried the MOI from 1 to 55 previously. After antibiotic selection for 6 days, all non-transduced cells were dead. I...
23 February 2017 281 5 View
I need to check transduction efficiency by using fluorescent microscope in the other building, however, I do not want my cells die during the process. A colleague doing research on immune cells...
14 February 2017 4,478 7 View
I'm trying to isolate RNA from suspended adipocyte now. The problem is that after centrifuging, the cells were still floating at the bottom, so when I remove all the media before adding RLT...
14 November 2016 2,324 4 View
I recently did chromatin extraction from brain nuclei samples. Since the individual sample amount is too small, I pooled 10 samples together (~40 mg). When I used the Epigentek Chromatin...
01 January 1970 321 7 View
