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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...
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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,...
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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....
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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...
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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...
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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...
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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...
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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...
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