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Questions related from Glen Drake
I want to use a RALA peptide vector, and I have found that some studies centrifuge the plasmid/RALA complexes and resuspend the pellet before use, while others seem not to do so. It seems like the...
06 June 2023 2,560 2 View
The Kozak sequence is 5'-gccRccAUGG-3', but the only amino acids that can be coded for by codons that start with G are valine, alanine, aspartic acid, glutamic acid, and glycine, so how is...
09 March 2023 8,250 1 View
I'm working with a non-model organism, and have recently come to understand that different terminator sequences have significant impact on RNA localization and translation efficiency, so I want to...
23 February 2023 3,268 0 View
I want to use secreted bioluminescence to identify successfully transfected cells, however that would require each individual cell is in its own little isolated well so that the successfully...
22 February 2023 7,669 1 View
I'm trying to make a stable transgenic line of a rather delicate eukaryote, so I want to activate transgene expression inducibly, but that means I need to express some kind of intermediary protein...
07 February 2023 7,467 0 View
I have been under the impression a Kozak is just GCCACC preceding the start codon, but just learned it also includes a G just after the start codon, i.e. GCCACC ATG G. If I am encoding a peptide...
06 February 2023 6,544 0 View
I have been thinking that I will just use EGFP to know whether my attempted transfection was successful, but I've been coming across some publications that say over expression of GFP can cause...
22 January 2023 4,287 2 View
Benchling's tutorial for automatic generation of primers for gibson assembly, https://help.benchling.com/hc/en-us/articles/9684273545997-Clone-using-the-Gibson-Assembly-Wizard , states that "Note:...
17 January 2023 6,929 0 View
I'm new to using CRISPR, and all the protocols I'm reading are just telling me to order the oligos for the primers for gRNA synthesis. Is this what everyone does, or is there a way to feasibly...
15 January 2023 7,732 3 View
Can I use PEI to simultaneously deliver a CRISPR RNP and the plasmid? Obviously PEI can package and deliver the plasmid, but I am having trouble finding information about PEI's ability to package...
11 January 2023 5,731 1 View
I have an idea for an organism i want to make, but to produce them i will likely need a vector that is no more than about 100 nm in diameter. I have been considering using BSA-PEI particles, but i...
11 January 2023 350 0 View
I found this study, https://www.biorxiv.org/content/10.1101/248179v3.full, and I really want to use the technique, but I'm concerned that monoavidin may have trouble binding a large DNA fragment...
18 September 2022 7,321 0 View
I'm looking at this study, https://www.biorxiv.org/content/10.1101/248179v3.full , and it uses a monoavidin - biotinylated donor DNA link to create a fusion of the donor DNA and the RNP so that...
18 September 2022 6,637 0 View
This was my original plan, however I was just reading this review A review of emerging physical transfection methods for CRISP... and it states that "When using lipofection, the cargo only...
14 September 2022 5,132 1 View
Hi, I'm trying to measure the relative abundances of complex II of the ETC with a western blot. I know PVDF is best for low abundance proteins and nitrocellulose is best for low background noise,...
04 April 2019 4,050 4 View
Hi! I'm looking at this study Mice Fed Rapamycin Have an Increase in Lifespan Associated w... , and their excel microarray data has negative log2 values, which doesn't make sense to me. Can...
04 April 2019 5,373 4 View
Is there a limit on the size of DNA fragment that can be bound by a biotin-monoavidin bond?
01 January 1970 496 1 View
