I want to add AviTag with my protein at N and C terminal and to express it in mammalian cell line.
My cloned gene contain a signal sequence at N terminal . How can I check the nucleotide sequence is a correct signal sequence. The following uppercase letter are signal sequence cloned in...
01 February 2019 7,668 2 View
I am planing to get two gene together for cloning purpose. My plan is first amplify the gene "x" using a set of primer which will amplify gene x without stop codon. I will amplify another gene "y"...
08 September 2018 6,732 1 View
Can I preserve my Transfected HEK-293 Cells IN -80 degree for few days, Later these will be being used for Microsome Isolation?
08 September 2018 8,593 1 View
My construct contains Htt 96Q followed by BirA* with a 6 amino acid linker between them. Than what will be size of my expressed protein if molecular mass of BirA* is 35.2 KDa? Based on amino...
07 August 2018 3,720 1 View
I am using knockout HEK 293 cell.
06 July 2018 4,511 2 View
Why I found colony for negative control on selection plate (contain only double digested vector : one blunt (by EcoRV) and one sticky (by SalI) end with ligation mixture, no insert, after...
06 July 2018 363 1 View
I want to ligate my 1kb insert (PCR product) with 6 kb vector, both are digested by ECORV (creating blunt end) and Sal1 (creating sticky end). How? Is blunt end may create problem in cloning?
05 June 2018 6,835 3 View
I am using Polyst. Flat bot. high Bind. MP blank (Costar) micro-plate for making ELISA (HIV ), when i am using this blank plate for ELISA assay , it is found reactive with positive control while...
31 December 2016 5,797 6 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 2,824 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 8,106 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 641 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
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
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 3,851 1 View
Hello, if you made a transwell assay where you incubate the cells with a nanoparticle-encapsulated drug and considering that in the oposite compartment you'll have both the free and encapsulte...
03 August 2024 2,730 1 View