I'm interested to learn about the times, efficiency, repeatability etc.
We'd like to produce recombinant proteins using a vector with His-tag in N-terminus. Our gene is inserted after 10-15bp after the His-tag. At the final step of purification and cleavage, how can...
31 December 2015 3,811 9 View
I've submitted a research article that deals with new biomarkers, and is based on samples received from cancer patients. The patients didn't give the blood for this project, however they consented...
09 October 2014 7,150 14 View
I've beem asked to convert some equations from Word to MathType. Unfortunately, I've not the software. Is there anybody that can help? Are there any online tools for such convertions?
08 September 2014 7,028 1 View
I'd like to perform a stable transfection to human cancer cell lines. I performed the transient with pCVMTNT vector, but I want a stable expression of a protein, which I study. The cell lines I'm...
08 September 2014 2,828 5 View
I'd like to study human resistance to alkaloids of the vinca cell line.
07 August 2014 1,337 4 View
After the Nick Translation protocol (abbottmolecular) of my PCR product (325bp product), the electrophoresis data are not good. There have been tested two different periods of time as well two...
05 June 2014 5,867 9 View
I'd like to perform some experiments by applying metronomic chemotherapy with 5FU and Oxaliplatin in colorectal cancer cell lines. Has anyone used a particular concentration for this application?
02 March 2014 1,099 2 View
I'd like to design a complete protocol for expressing protein in mammalians by using plasmid expression vectors. However, I've never dealt with culture of competent cells, so I'm not familiar with...
02 March 2014 7,190 5 View
I'd like to design a probe for FISH. The area is non-coding, but the sequence is known. The size of the probe should be 500-1500bp. Do I need to use DOP PCR with the desirable primer? Does anyone...
31 December 2013 632 6 View
We would like to analyze the data from antibody array after scanning.
11 December 2013 9,086 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
After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...
25 February 2021 5,712 3 View