Hi,
I wanted to do bisulfite treatment on tRNA (length 72bp) and design taqman probes for methylation specific PCR. Have anyone done this before and could you please let me know the things which i have to take care of while designing the probes?
Hi, I am looking for a detailed protocol where I can measure both mitochondrial and cytosolic calcium dynamics using live cell imaging? The buffer conditions and the optimal ionophore...
11 December 2017 5,545 0 View
I need a very good protocol for competition binding assay. I have a complex and a third protein. I expect the third protein to dissociate the complex by binding with one partner of the complex....
10 November 2014 5,936 3 View
I am trying to purify two proteins. Protein 1, is around 16kd, sequence verified, expression verified via western blotting. problem is induction. it is not induced. i tried various concentrations...
09 October 2014 2,531 10 View
I am trying to subclone gene of my interest along with tag from pcdna3.1 to pET28b vector. As the insert has its own ATG, gene of interest and fusion tag in frame, I want to cut the whole lot and...
08 September 2014 551 6 View
See above.
05 June 2014 3,508 8 View
Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...
03 March 2021 687 1 View
Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...
02 March 2021 4,029 3 View
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
To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...
01 March 2021 8,683 4 View
Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?
01 March 2021 5,311 1 View
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
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
Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...
26 February 2021 4,777 4 View
Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...
26 February 2021 5,029 2 View
Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...
25 February 2021 6,969 3 View