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 this to normalize my input DNA in the reaction.
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...
04 May 2018 3,896 6 View
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...
03 April 2016 976 3 View
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...
04 May 2014 5,745 4 View
Assuming the use of FCS+10% DMSO as cryoprotectant and 1ml of final volume.
07 August 2013 9,478 7 View
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...
09 October 2012 9,186 5 View
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...
31 December 2011 7,088 5 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
This larva was captured using a plankton net in the Persian Gulf during the summer. I believe it may be a Facetotecta nauplius.
08 August 2024 3,746 4 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you
08 August 2024 4,733 2 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...
07 August 2024 6,034 3 View
Hi all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...
06 August 2024 5,801 4 View
There are a huge number of methods for studying objects in space, according to the senses (and not only). Mechanical, thermal, optical, acoustic, electrical, magnetic, based on particle beams,...
06 August 2024 7,102 0 View
During low-temperature testing, new diffraction peaks that appear could be indicative of several phenomena. In one of our tests, we observed notable new peaks around 40° and 45° in a specific...
06 August 2024 726 3 View