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Questions related from Celina São José
We performed 4C-seq according to the van de Werken protocol. In order to sequence the 4C libraries produced in Hi-seqX Illumina platform, we need to design long primers with P5 and P7 adaptors, as...
26 February 2020 3,798 0 View
I have a cell line that stably expresses Cas9. However, I am not sure how to deliver the sgRNAs or gRNAs. Is it possible to buy tracRNA and ligate with sgRNA oligos before transfection? Or can I...
07 May 2019 3,532 4 View
05 May 2019 6,439 5 View
I am using Dan Swarr and Dave Frank's protocol to ligate oligos to the PX459 vector, using Stbl3 cells (invitrogen). However, I get very few colonies (max 3) after transformation using 20ul of...
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I am culturing primary cells and use DMEM F12 (1:1) cell culture medium supplemented with human insulin and EGF. How long are these components stable for in the medium? Thank you for your gentle...
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I want to perform 4C-seq on adherent malignant cells. However, after fixing and before lysis my cells don't have cytoplasm anymore and the DNA is fragmented (observed by a smear in a gel). Did...
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I want to isolate stomach epithelial cells, from both human and mouse, for primary culture. Does anyone has an easy to follow protocol withou resourcing to 3D cell culture?
10 October 2018 4,238 1 View
I need to transport embryonic organs fixed in 4% PFA and previously stained in b-gal solution (LacZ) to another country. Our objective is to embeed the tissues in paraffin. Is 4% PFA the best...
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