Kristen Navarro

10 Questions 2 Answers 0 Followers

Questions related from Kristen Navarro

How do I create forward and reverse gRNAs for dCas9 CRISPR?

Hello everyone! I am reading a protocol for CRISPR using a dCas9, and I came across the following passage in the gRNA section: A. Design sgRNA oligos Fwd (all): ACCGNNNNNNNNNNNNNNNNNNN Rev (Sp):...

21 August 2023 9,047 0 View

First time doing lentiviral transduction - how much antibiotic should I add to selection media?

Hello! I am performing a lentiviral transduction on human fibroblasts for the very first time. The vector I am trying to insert contains puromycin and ampicillin resistance. How much ampicillin...

07 August 2023 5,479 4 View

Best primers for qPCR control?

Hi! I am trying to optimize a qPCR experiment using mouse tissues. I am trying to use a GAPDH control, but I think my primers may not have been designed well. Does anyone have primers for GAPDH...

18 July 2023 5,887 3 View

What is a good western blot control that is below 30 kDa?

Hello! I am trying to determine the best controls for my western experiment. I was wondering what the best controls are with a small kDa size?

07 July 2023 566 3 View

How do you make 25 mM HEPES buffer?

Hello! I need help making a 25 mM HEPES buffer with a pH of 7.4. I've tried calculating the dilution, but I keep messing up. The stock solution I am planning to use is: 30 mM HEPES Buffered...

05 June 2023 7,769 1 View

What is the best plasmid & protocol for live cell imaging? I have never done this before?

My graduate work was in C. elegans, and this is my very first time performing live cell imaging in human cell lines. (fibroblasts) I would like to use live cell imaging to track two proteins of...

19 May 2023 1,082 2 View

Best software for Western blot band quantification?

Hello everyone! I'm currently having some trouble looking for software that can view and quantify Western blot bands. Does anyone have any recommendations? I am currently using a MacBook Pro for...

24 March 2023 663 4 View

How can I overexpress a gene of interest using a lentiviral system?

Hello! I am trying to find a plasmid and protocol to overexpress a specific gene of interest in fibroblasts. Does anyone have any recommendations?

24 March 2023 2,858 0 View

Is there any software/web tools that can convert cDNA to RNA?

Hello! The sequences I am examining in the Ensembl Database are in cDNA format only, but I need to look at the RNA sequence. How can I convert the cDNA sequence into RNA? Most online tools only do...

09 February 2023 1,187 3 View

How can I generate a PDB file for a new, uncharacterized amino acid sequence?

I'm trying to use EzMol to illustrate a protein sequence, but there are no PDB files for the sequence I'm trying to analyze. (NCBI: XP_006501616.1) How can I create a PDB file?

08 February 2023 7,805 3 View