Leonardo Chicaybam

7 Questions 25 Answers 0 Followers

Questions related from Leonardo Chicaybam

Can someone share a primer/probe set that can amplify the same locus in genomic DNA from mouse and human?

I have a DNA sample that contain mixed mouse and human cells. I need a set of primer/probe for Taqman that can amplify with the same efficiency a locus in mouse and human genomic DNA, so I can use...

07 July 2018 946 0 View

How do I evaluate transfection/electroporation efficiency after only 4h?

My experiment consists of electroporation of PBMCs (peripheral blood mononuclear cells) with plasmid DNA and use of modified cells 4h after electroporation. However, after 4h I still won´t have...

05 May 2018 3,863 6 View

It is possible to sort cytokine producing cells, isolate DNA and perform PCR?

I would like to stain lymphocytes using an anti-IL2 antibody (for example), sort the positive cells using a flow cytometer and isolate DNA for PCR. Is this possible? Do you have some reference...

04 April 2016 954 3 View

Trouble in culturing LCLs. Can anyone help?

I am having trouble with my cultures of LCLs. After thawing, the cells are ok and they proliferate for 1-2 days. But after this initial proliferation the cells start to die and don´t recover. The...

05 May 2014 5,714 4 View

What is the maximum number of PBMCs that can be frozen in one vial?

Assuming the use of FCS+10% DMSO as cryoprotectant and 1ml of final volume.

08 August 2013 9,442 7 View

What is the best way to overexpress miRNAs?

I want to use the pLL3.7 lentiviral vector (U6 promoter) to overexpress miRNAs. However, I have some doubts regarding the sequence that I have to clone in the vector to achieve overexpression. I...

10 October 2012 9,153 5 View

Viability of murine lymphocytes.

I am isolating lymphocytes from lymph nodes of C56BL/6 mice. After the isolation I activate them with 4ug/ml of soluble anti-CD3 (clone 2C11) and 1ug of soluble anti-CD28 (clone 37.51) and 24h...

01 January 2012 7,056 5 View