Is there a way to predict a dimer formation of fused proteins?

We have got a recombinant protein which consist of target molecule and EGFP. However, the Mw of this fused protein is two times bigger than it was predicted. We are wondering if it could be a...

09 October 2015 7,333 12 View

How can I get rid off DNA for following with RT-qPCR?

We would run RT-qPCR to measure viral gene expression profile. In order to extract RNA from infected cell line Qiagen RNAeasy mini kit was used. However, big enough amount of DNA was still...

03 April 2015 5,803 15 View

What is the average yield of protein of interest express in mammalian cell line under CMV promoter control after transfection?

09 October 2014 5,660 9 View

Can EGFP fused proteins be expressed separately?

We have fused a protein of interest with EGFP into plasmid vector under CMV promoter control. In WB we have got two bands correspond to EGFP itself and fused recombinant protein according to Mw....

06 July 2014 400 9 View

Does anyone have an experience in 3' end viral genome amplification?

We have an issue to amplify 3' end of dsDNA virus genome (ASFV). Basically, we use Long-Range PCR to amplify whole viral genome. Everything was fine except 3'end region. We still have around 6 kb...

03 April 2014 6,429 16 View

What is a good way to estimate virus titer?

There is no doubt that widely-used Kaerber or Spearman or other methods based on virus back titration using 10- fold dilutions have shown acceptable results. Anyway, if we need to estimate virus...

06 July 2013 1,486 13 View

Does anybody know how to predict antibody binding sites on protein molecules?

Currently working on glycoprotein structure (CD2v) in frame of prediction modeling. Given glycoprotein has a lot of glycosylation sites and our question is could the glycosylation affect Ab...

11 December 2012 7,560 2 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

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 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

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

1 nm thick MgO thin-film to be deposited on Si wafer using thermal evaporation. What will be the optimum deposition conditions?

Favourable conditions of base pressure, rate of deposition. Also, by which characterisation technique to be used to verify the thickness of the deposited film.

28 February 2021 3,640 3 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

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View