Bruno Salomone Gonzalez de Castejon

10 Questions 37 Answers 0 Followers

Questions related from Bruno Salomone Gonzalez de Castejon

Discrepancies in N/P Ratio When Using Molar vs. Atomic Count of Nitrogen and Phosphorus?

Hi, I am calculating the N/P ratio to encapsulate dsDNA into a LNP. I am using two different methods, but obtaining a worrying difference in the result. I believe it could be a mistake in one of...

28 December 2024 5,821 3 View

Optimising pegRNAs or sgRNAs for prime and base editing?

hey, im designing an experiment to optimise prime and base editing epeg/sg RNAs based on their efficiency and off target effects. I understand that using a cell line easy to transfect such as...

16 October 2024 6,813 2 View

Why are my nanoliposomes turning purple??

Hello, I am making lipid nanoparticles using the thin film hydration method. Formula is DODMA:DSPC:Cholesterol:DSG-PEG2000 at 60:20:17.5:2.5 %mM in 5mM total. After hydration in sodium acetate...

17 June 2024 7,237 6 View

Isssues making a thin film of lipids for nanoliposomes?

Hello, Im trying to make a nanoliposome to encapsulate plasmid DNA. I am using the standard thin film method followed by resuspension in sodium acetate 25mM pH4 with my DNA (Im using the...

06 June 2024 3,802 3 View

Performing restriction digest on total DNA isolation from mammalian cells?

Hello, I am transfecting linear DNA along with an Adenovirus transduction to HEK cells and need to isolate the DNA from both the linearised plasmid and viral genome at different passages for...

14 November 2023 4,904 3 View

Will performing a PCR on a multimeric plasmid population yield multiple bands?

Hello, I have performed some recombineering protocols and realised that the chances of my plasmid being in a multimeric state are quite high. I previously designed 7 primer pairs that will...

19 March 2023 4,097 4 View

Reecombineering - why are my positive recombineered clones not growing in LB broth?

Hi All, I am currently doing recombineering to insert two mutations on a plasmid, but after successfully recombining the sequence, the cells from my plates don't grow on LB broth. This is my...

25 February 2023 9,004 4 View

Anyone with Red/ET Recombineering experience?

Hello! I am carrying out, for the first time, some recombineering experiments, and any tips or advice not apparent from current protocols that you may have from experience would be really...

15 December 2022 3,516 3 View

Does anyone know of a DNA stuffer sequence?

Hello, I am looking for a 3kb (approximate) stuffer sequence, I have seen that people use 'non-contiguous lambda bacteriophage DNA segments with no intact coding region' but I'm not too sure what...

30 November 2022 487 0 View

Anyone with experience in vector design with insulators?

Hi All, I am designing a vector carrying a bicistronic transgene with two promoters that will be integrated into the host's genome. I am thinking of adding three insulators (the same type): two...

17 November 2022 1,925 1 View