To synthesize chimeric receptor, should I used real gene sequence or artificial sequence?

07 February 2019 1 2K Report

I'm synthesizing a chimeric antigen receptor (CAR), and I have the whole sequence of the amino acid.

Now the problem is, to ask company to synthesize the DNA of the CAR, should I send real gene sequence or artificial DNA sequence predicted by computer based on the amino acid?

Pavel Grinkevich

It depends on what you're planning to do with it afterwards. If you're going to express it in a particular host, for example E. coli, you can optimize the codons for that host.

More Suipwksiow Wjkp's questions See All

Can FDA-approved drugs be used in other disease setting?

Hi Everyone, I'm exploring the anti-tumor effect of some drugs. I'd like to test some FDA-approved drugs which are not used to treat cancer. My question is, if there is any anti-tumor effect for...

14 December 2020 1,464 1 View

How to know if my protein forms a dimer or not?

Hi everyone, The protein I studied is a membrane protein. Some study indicates that there is possibility that this protein can form a dimer. I did standard western blot for this protein and my...

20 October 2020 6,935 7 View

How to ensure a marker antibody won't activate T cells and affect the result?

I'm recently buying some anti-CD3 and anti-CD28 to activate T cells. I also notice some of this clones have been conjugated with fluorescence for the purpose of labeling T cells for flow...

06 October 2019 7,129 3 View

Why my gene in single-cell sequencing showed zero value?

I have many single-cell sequencing data. I checked my gene of interest in these data. However, all of these single-cell sequencing data showed that my gene has no expression, no matter in immune...

03 January 2019 1,707 3 View

Why can't I detect the surface protein by western blot?

I was doing an over-expression experiment of a gene for a year. It is a surface protein. The biggest trouble is that I can never detect the gene overexpression by western blot. Interestingly, when...

29 November 2018 6,311 3 View

For CRISPR, transfection or transduction?

For CRISPR, transfection or transduction? The CRISPR vectors I bought is lenti vector, so I can either do transfection or transduction. But which is better??

13 March 2018 4,447 3 View

Can I use NEB mutagenesis- substitution method to clone sgRNA into the vector?

Can I use NEB mutagenesis -- substitution method to clone sgRNA into the vector? I felt it's much easier. Just design the primers containing the sgRNA bp, conduct PCR and do a ligation. And most...

08 March 2018 4,991 1 View

What's the best way to eliminate Mycoplasma?

Recently check the cell lines, 60% of the cell lines are infected with Mycoplasma. Some common antibiotics for treating Mycoplasma were used to treat cells for 3 weeks, but mycoplasma is still...

14 February 2018 9,511 2 View

Why using RecA- bacteria strain?

The growth instruction for a CRISPR plasmid said that the plasmid need RecA negative E.coli strain like Stbl3 to grow. What is special for RecA negative strain? Why the clone need RecA negative??

19 January 2018 5,047 6 View

How is heavy chain and light chain of an antibody connected?

I have CHO cells that transduced with two vectors expressing light chain and heavy chain of an antibody respectively. How do they automatically associate with each other and form antibody? What...

27 December 2017 5,668 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

What solvent can I used for Paal Knorr reaction between 6-amino hexanoic acid and 2.5 hexanedione?

I am try to make the Paal Knorr reaction between 2.5 hexnedione and 6-amino hexanoic acid, my problem is type of solvent to use, because 6-hexaminohexanoic acid is soluble in water but I am not...

02 March 2021 4,443 2 View

Ester cleavage conditions?

Hi all, I need to cleave the ester in Ethyl 2-amino-1H-imidazole-5-carboxylate to make the carboxylic acid and retrieve the product from solution to use in downstream applications of linking to...

01 March 2021 8,766 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

What is the most common number of epitopes in an multi-epitope Vaccine?

hi everyone, I'm interested in the field of vaccine design specifically multi-epitope cancer vaccines , I want to know if there is a minimum and maximum number of epitopes we could use in a...

27 February 2021 3,882 3 View