After purifying protein of interest their is non-specific multiple bands above and below the protein of interest band. Can DEAE method be used?

I am trying to purify a protein and after purification the expression of protein of interest is pretty good but their are non-specific bands below and above the protein of interest. Can anyone...

08 September 2015 2,957 12 View

I had done PCR standardization to amplify my gene of interest of 1.5kb from cDNA. When I repeated the same protocol, no amplification was seen. Why?

I had done PCR standardization using cDNA as template. I did too many pcrs and finally standardized to amplify the gene of interest of 1.5kb from cDNA. I amplified 2 different genes but of same...

10 November 2014 2,919 8 View

I did cloning for phusion protein. I did westerns and getting a band of interest along with a band with low molecular weight, why?

I had cloned my gene of interest in to a vector, which has fluorescent protein tag. I did restriction digestion and did insert release of 1.4kb and fragment release having both gene of interest...

06 July 2014 6,175 8 View

Why am I not able to get a clone of gene tagged with GFP?

As previously discussed, I redesigned the primers. I tried triple ligation, it did not work. So, the gene of interest was restriction digested from the vector containing gene and was amplified...

31 December 2013 604 9 View

I am not getting a desired amplification for new set of gene specific primers designed?

Forward primer: 79.6 degrees melting temperature and Reverse primer: 81.1 degrees. Reverse primer has strong secondary structure and no dimer. I tried pcrs setting with high denaturation temp and...

10 November 2013 2,138 16 View

How to write scientific paper related to clinical psychology in neurological disorders?

I have collected the data and analysed. But writing discussion is little difficult, as I am looking for pathology in brain for psychological symptoms. Please suggest me as to how the discussion...

08 September 2013 9,551 5 View

PCR reaction is not working and not showing any amplifications, while positive control shows. I changed buffer, Mgcl2, dNTPs mix, water. Why is this?

I have been doing genotyping for a mouse model. I isolate the genomic DNA from Alzheimer's mice tails. I have done many PCRs, but this batch of isolated genomic DNA, has good concentration and...

08 September 2013 9,497 49 View

Can I use a combination of acids like Hcl and H2so4 / Hno3 in adjusting the pH of Tris buffer saline?

Tris buffer saline of pH-7.4. As in case of emergency like one of the above mentioned acid gets over while adjusting the pH, than rest of the pH could be adjusted using other alternative acid. Is...

06 July 2013 7,655 2 View

In PCR, why do the salts differ for particular polymerase enzymes like Mgcl2 for Taq and MgSo4 for Kod polymerase?

However both salts have Mg ions.

05 June 2013 7,162 2 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

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02 March 2021 3,060 3 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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