I need help on the most appropriate transfection approach and conditions to optimize cell transfection when establishing stable cell lines expressing any of the cervids' prion protein genes and their polymorphisms.
I am trying to check the polymorphisms of a particular gene from human DNA samples. A set of primer in a published publication allowed the amplification of the DNA hence permitting the analysis of...
02 March 2019 8,427 5 View
I subjected some genomic DNAs to PCR reaction and later sent then for sequencing to analysis for polymorphisms in my gene of interest? Some of my sequence results came out fine while some came out...
01 February 2019 9,724 7 View
I carried out a deglycosylation test to see if there will be difference in the electrophoretic mobility of C57BL/6 infected-ME7 and SShpPrP Tg ( transgenic mice expressing sheep prion protein)...
01 February 2019 6,065 2 View
I am trying to establish a stable cell line expressing a gene of my choice using a prion protein knock out cell line, Lipofectamine and Zeocin as selection antibiotics. Not ruling out the fact...
11 December 2016 9,133 7 View
I did tail biopsy to genotype a particular gene. The result came out positive for my desired gene but two days after using same lysed genomic DNA, it came out negative. In my confused state, l...
11 December 2016 5,779 7 View
6H4 monoclonal antibody known to be effective in capturing cervids Prion Proteins has been discontinued by Prionics. I need suggestions on antibodies either monoclonal or polyclonal that could be...
11 December 2016 2,427 2 View
I need the procedure of how to establish a stable cell line urgently.
07 August 2016 6,682 2 View
Just want to know the volume of lysis buffer to added to weighed cell pellet or tissue during cell lysis.
06 July 2016 4,699 0 View
I am trying to establish a cell line using Zpl cell line- PrP knock out and LTX/PLUS lipofectamine reagent as transfection reagent.
06 July 2016 426 4 View
I need protocols or procedures for extracting Mi RNA from tissues in experimental animals.
04 May 2016 1,262 4 View
I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...
09 August 2024 9,621 0 View
Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...
09 August 2024 2,822 3 View
I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...
07 August 2024 7,535 4 View
Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...
07 August 2024 8,105 4 View
Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...
06 August 2024 639 2 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,987 3 View
I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...
04 August 2024 1,200 2 View
I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...
04 August 2024 9,913 7 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 would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...
03 August 2024 3,850 1 View