Why do I receive completely different sequencing result?

Dear all, I cloned 25bp fragment into a vector. My no-insert ligation has no colonies and my insert+vector ligation has many. Which is the first clue that I have cloned the fragment. Than I...

11 December 2019 7,539 8 View

How can I eliminate yeast contamination?

Dear friends, We kindly get U-251 MG cell line as a gift from an other lab. after 3 passage we have determined that cell line has yeast contamination. Each stock is contaminated while other...

11 December 2018 5,313 8 View

How Can I detect artificial miRNA in stable cell lines?

Dear all, I have generated Stable Cell lines via G418 selection. I have used pCMV-miR vector.Except Neo-GFP region, my chance is 2/5 to integrate genome without damaging CMVpromotor-insert-PolyA...

07 August 2018 8,094 2 View

Is it possible to clone without PolyA signal?

hey, I have generated stable cell lines with random entegration of pCMV-miR vector. Vector contains a neoR gene fused with GFP with IRES. I got some GFP+ and GFP- colonies. and GFP- colonies...

07 August 2018 612 3 View

G418 selection cell death problem?

Dear Friends, I performed a kill-curve experiment and determinated that ; at 400mg/ml g418 consantration only 5-10 non-profilable cell survived at 7 day in 48 well-plate, plated with 50.000 U87...

05 June 2018 7,987 3 View

During luciferase assay my miRNA target luciferase gene's coding sequence?

Hey, I have a small problem. I have cloned my target gene's UTR into pmir-Glo vector. But the problem is that frefly luciferase gene also contains miRNA target sequence. Thus when I clone my UTR...

04 May 2018 9,073 0 View

Transfection efficiency as well as transgene expression in high passaged GBM cell lines?

Dear all, I am working with U-87 MG (a glioblastoma cell line whose origine is unknown but male) and T-98G. I have clonned my miRNA genes into pCMV-miR vector that express transgene via CMV...

03 April 2018 6,728 3 View

Does anyone have home-made buffer for Amaxa electroporation u-87 cell line?

Hey friends, I am working with U-87 cell line. Standart transfection reagents show no transfection efficiency on U-87 cells. We have Amaxa nucleofector but its reagents are highly...

03 April 2018 9,608 0 View

How to eliminate low transfection efficiency?

I am working with glioblastoma U-87 cell lines. I have pCMV-miR expression vector ~8kb. I tired both Transfast, Fugene HD, PEI, Jetprime. They had max %30 efficiency. Transfast best with 1:1...

03 April 2018 9,602 8 View

Can 1 kb genomic fragment be used as pri-miRNA?

Hey all, To overexpress a miRNA; instead of pre-miRNA sequences, pri-miRNA sequences are also being used driven by RNAP II promoters. in articles these fragments nearly miRNA+100-300 bp flanking...

03 April 2018 3,764 5 View

Does the pressure(-600 mmHg) affects pH?

When I conducted the degassing experiment using decompression under anaerobic condition(and I used ferrous based chelating agent), the oxidized ferric were observerd. So, could you tell me the...

03 March 2021 4,892 5 View

Chlorinated organic formation in odor control scrubbers using hypochlorite?

I'm involved in a study of odor control technologies for municipal wastewater treatment plant. One of the control options involves a chemical 2-stage (acid/alkaline) packed bed scrubber. The...

03 March 2021 3,661 2 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 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

How to make fractions of chloroform extract of plant, using seperating funnel?

28 February 2021 8,466 2 View

I want to performe MALDI-TOF MS for bacterial identification, I would appreciate it if anyone can tell me the matrix solution preparation protocol ?

based on what I've studied on multiple papers, for peptide and protein identification Alpha-cyano-4-hydroxycinnamic acid (CHCA) is an appropriate matrix to use, I also know that the CHCA powder...

26 February 2021 6,799 3 View

Which database would give me this list of chemical components?

Dear colleagues: I would like to know if anyone knows of a database that gives me the parameters pH, K, K, Mg, P, C, N, Zn, Cu, Ni and Cd of common foods. I find a lot of information of proteins,...

26 February 2021 5,525 3 View

How can we theoretically calculate Elemental analysis(percentage) of atoms in covalent organic framework?

For example in one research article the Elemental analysis values of COF-TpPa-1 ( Triformylphloroglucinol (Tp) (63 mg, 0.3 mmol), Paraphenylenediamine (Pa-1) (48 mg, 0.45 mmol) are as follows...

24 February 2021 1,265 6 View

How to Control Whiteflies in coconut leaves?

24 February 2021 1,005 1 View