Hello,
Do you have beta actin primer sequences for qPCR? I will study with 3T3 (mouse fibroblast), Raji (Human B cell), U937 (Human monocyte), HEK293 (Human embryonic kidney). May you suggest sequences your worked?
Thanks...
Best regards...
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Hi, I will do transfection with fugene but, I tried to enter three plasmids in cell. There is no selection part of plasmids, and I will try to plasmids enter to in cell one by one. How to do...
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Hi, I did western and you can see my beta actin bands. I put 10 uL primary antibody for my interesting protein, but my professors said 25 uL should be. I donot understand. Wouldn't I have seen a...
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Hello, I have band of my negative control. I am sure there is no contamination. I used new everything, sterile and change my gloves 5 times. I did negative control mix separately my samples. My...
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Hello, Last week I asked my PCR results and I send Sanger seq. Well, there is one colony including mutation, I guess. How to cleaning my sample for Sanger? The Sanger result belongs to the second...
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Hi everyone, First of all, thank you for attention and answers. I did reverse transcriptase PCR for obtain cDNA from RNA. I used 3 different cell line RNA and my results looks fine....
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Hiii, I try to make mutations with QuikChange II XL Site-Directed Mutagenesis Kit. When I did first mutation, I got colonies and also mutation was succesfull. However, although I tried everything...
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Hi, I have a plasmid and designed site-directed mutagenesis primers, but my professor wants from me check plasmid with GFP after transformation? I donot know how to do that. Because our plasmid...
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Hi, I have 2 specific variants and i want to create mutatation with mutagenesis primers. How can I design these primers? I found QuickChange Primer Design Tool but I couldnot use. Do you have any...
23 October 2021 5,620 3 View
Hi, Do you have any CRISPR/Cas9 system tool such as in silico PCR? I designed a crispr vector, before order it, I want to check. Thanks.
12 September 2021 3,069 0 View
Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...
04 August 2024 7,282 2 View
I am conducting my research regarding blood coagulation in patients with CKD. I observed a reduction in post hemodialysis aPTT and PT values compared to pre hemodialysis values. But most of...
30 July 2024 3,467 0 View
Dear colleagues and scientists: I am asking for your help and insights to develop and explore the feasibility of using an immortalized kidney cell line, for example, HEK293T, for the induction of...
29 July 2024 5,622 3 View
I'm new to primary human fibroblast culture and I'm studying aging using cells from old and young donors. Some research groups have used media supplemented with FGF2, so I cultured cells in the...
29 July 2024 3,030 2 View
hi every one I am making vector construction (for fusion proteins) and in this moment I wanna to amplification of ADAM17 prodomain with PCR. to yet, I couldn't amplified the ADAM17 prodomain with...
21 July 2024 8,660 1 View
Hi, I need help in detaching adherent monocytes from well plates. I isolated the PBMCs from buffy coat and I did CD14+ positive selection to collect only the monocytes. I let them adhere on my...
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I am using fibroblast growth factor, purchased in powder form. What suitable substance should be added to get it in a liquid form? Thanks
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Hello, I have been performing Western Blots to study the protein levels of a rodent skin and muscle sample. However, I have not been able to produce homogenous reference bands. I suspect that...
26 June 2024 5,913 3 View
Hello, everyone. I would like ask one question. I transfected fibroblasts with EF1A-myod-puro using lipofectamine 3000. Promoter for puro is CMV. After transfection 48 hours, I did puromycin...
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Bfgf was aliquoted in media and transported at 22 degee Celsius. How long it is stable
15 June 2024 3,010 2 View