Can anyone help with gene replacement in a tumor cell line?

11 April 2014 1 9K Report

I am attempting to track a protein using a modified RFP. I must create a fusion protein but the protein must be expressed as a replacement for the original gene in order to understand the dynamics of expression under this specific promoter.

I would like to "tack on" the RFP at the end of the gene sequence. Any ideas?

Badges
Science topic

More Mitchell Duffy's questions See All

Can 5-FOA Diffuse Through Agar?

I am preparing to do a gene knockout in the filamentous fungi Coccidioides posadasii targeting pyrG/URA3 so that 5-FOA can be used for selection pressure instead of traditional antibiotic...

16 February 2021 169 4 View

Are there any simple materials besides activated carbon that can remove a variety of dissolved organic chemicals in water samples?

I am looking for a method to remove a suite of dissolved organic compounds (e.g. BTEX, PAHs, PCBs, acetone/acetate/acetic acid, phthalates, phenols, glycols, biocides, surfactants...) that are...

12 February 2020 9,656 3 View

Difference between Resistive and Inductive Switching?

Both types of switching are used to characterise the transistor ON-time (tON), rise time (tr), fall time (tf), OFF-time (tOFF). What are the differences between the two characterisation methods...

26 August 2019 4,797 1 View

Is there a way to simultaneously visualize DNA and protein following electrophoresis?

I am currently optimizing the conjugation of a DNA oligo sequence to a DBCO activated antibody and am looking for a method which could allow me to simultaneously visualize the amount of DNA which...

15 May 2019 9,228 3 View

Can anyone point me to literature exploring WHY researchers study what they do?

I'm looking at this issue in relation to education research in Africa, but very interested in insights from other fields and contexts, too.

04 September 2018 6,158 8 View

Does the order of LysC, PNGase, and trypsin in proteomics sample prep matter?

I was under the impression that one of the benefits of LysC is that it is active under high urea concentrations (8M) retaining protein denaturation in hard to digest proteins (such as ECM). PNGase...

11 June 2018 7,849 2 View

Log or square root transformation?

Hello, I recently analysed count data (in this case the no of stems per tree) using a GLM. The analysis was carried out on both log transformed and square root transformed data. Although...

10 February 2017 8,300 7 View

Does anyone have experience with methods to overcome fluorescent overlap between Doxorubicin and mCherry?

I am looking to assess the metabolic control of breast epithelial cancer cells in cancer-associated fibroblasts (CAFs) and the subsequent effect of this on doxorubicin chemoresistance using an in...

22 January 2017 2,175 1 View

The answer to TPN hyperglycemia control.

11 December 2016 6,168 6 View

Best Method for Quantifying miRNA in Sperm Samples?

We have collected sperm samples and wish to measure miRNA content. What is the best method/kits to go with from extraction of RNA all the way to PCR?

15 November 2016 3,488 2 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 View

No title

02 March 2021 3,060 3 View

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View