22 September 2017 3 5K Report

I'd like to fuse 3 fragments into a vector, but they have RE sites. How to connect them together without adding extra space? I don't wanna additional a.a to be translated.

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

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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...

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

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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?

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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

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