Do very low concentration of non-specific bands in purified protein sample interefere in Circular dichroism results ?

I purified two ~ 55KDa proteins having C-terminal His-Tag. The concentration of both the proteins are ~ 40 uM. One is the wild type and the another is the mutant protein. The proteins always...

02 March 2018 6,505 7 View

Can anyone suggest me where I can avail the Stopped Flow instrumentation facility in India?

I need to do an experiment on enzyme kinetics by using Stopped Flow instrument. But due to lack of instrumentation facility I am unable to progress in my project. Can anyone suggest me where I can...

11 December 2017 8,283 0 View

Why the PCR product after restriction digestion is showing two bands?

01 February 2016 5,592 6 View

I am facing a problem in Protein Expression. Why am I getting 3 induced bands instead of one?

I cloned a gene in pET28a(+) vector and transformed in E. coli BL21 for expression. After IPTG induction and SDS PAGE analysis I am getting 3 bands that seem to be induced together instead of one...

05 June 2015 5,674 9 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Is there any published or unpublished statistics on dropout rate (Dose1 to Dose-2) for Covid-19 vaccination in India, US, UK or any other country?

Are there any published or unpublished statistics or papers on the dropout rate (Dose1 to Dose-2) for Covid-19 vaccination in India, USA, UK, or any other country where vaccination has started at...

28 February 2021 4,237 1 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

How to reduce static electricity of powder after lyophilization?

I am working on synthesizing and then purifying peptides using the SPPS system. After purification of the peptide, the peptide powder is obtained using a freeze dryer. When measuring the powder,...

22 February 2021 10,099 5 View

How to minimise the risk of being abused the profile of potential participants tagged in a social media post?

Dear colleagues, I got this one in one project I did ethics review. It's great to hear about your thoughts/experiences. "Doing an open recruitment online (i.e., via Facebook) requires public...

22 February 2021 3,322 3 View

How to get the vector of a specific vibration mode in phonon dispersion?

For example, I have this phonon dispersion(Fig. 1), and now I would like get vector of atoms which describes TA1 in P. I know v_sim can show vibrational animation of specific mode, but I do not...

20 February 2021 4,332 4 View