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Questions related from Panagiotis Apostolou
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...
01 January 2016 3,888 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...
10 October 2014 7,227 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?
09 September 2014 7,112 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...
09 September 2014 2,899 5 View
I'd like to study human resistance to alkaloids of the vinca cell line.
08 August 2014 1,421 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...
06 June 2014 5,952 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?
03 March 2014 1,174 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...
03 March 2014 7,266 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...
01 January 2014 711 6 View
We would like to analyze the data from antibody array after scanning.
12 December 2013 9,163 1 View
I'm using 18SrRNA and β-Actin as housekeeping genes in my qPCR experiments. GAPDH and HPRT1 have higher Ct. However in many samples (CTCs), the 18SrRNA seems to be more accurate.
09 September 2013 9,592 9 View
I'm using many kits as well as Trizol for extraction of RNA from cancer cell lines and cancer stem cells. In many cases, after the extraction, I get the 18S-28S bands but I don't get other genes...
06 June 2013 767 14 View
I'd like to find out the best molecule/substance for inhibition of RNase. There are many enzymes, but many are using also β-mercaptoethanol. I'd like it for my RNA extraction assays.
06 June 2013 7,961 4 View
I'm trying to insert a PCR product into a M13 vector, by using SmaI enzyme. After the Blund End repair, and the ligation, I do not see anything on agarose gel. Without Blund End Repair I see the...
06 June 2013 4,507 9 View
After digestion of M13m18RF DNA with SmaI, for 1 hour, I got the below picture. The digested product is smaller than the uncut. Could it be possible? Because in many photos, I have noticed that...
06 June 2013 8,978 1 View
I use siRNA molecules for knockdown assays in cell cultures. I use both molecules 19b lengh with dTdT overhang. I have tried also with dUdU. Can anyone tell me, the basic differences between all...
04 April 2013 4,356 1 View
I'm trying to create a cisplatin resistant cancer cell line, HCT-116. Does anyone know the optimal concentration that I must add to the culture? I started with 1μg/ml of CDDP. I noticed that the...
03 March 2013 457 32 View
I'd like to create a cell line from a tissue. Until now, I remove with trypsin the cells from the tissue and culture them. Also I keep the tissue pieces in petri dishes. In the culture I use RPMI...
03 March 2013 3,752 6 View
I have an unknown DNA product (about 500bp) which I want to sequence. I cannot insert it into a vector, because I have no vector. The company I do sequence for cannot do it only for one sample. Is...
03 March 2013 3,943 0 View
I'm interested to learn about the times, efficiency, repeatability etc.
02 February 2013 687 0 View
With TRIZOL method the RNA is degraded and we observed high absorbance in 230 (>2,5). The ratio is good enough (1.8-2.1).
01 January 2013 3,404 5 View
