54 Questions 56 Answers 0 Followers
Questions related from Mahbobeh zamani babgohari
Antioxidant assays(I.e. DPPH) are normally conducted on fresh tissues. How about freezing samples and do the assay after some time later. Thanks
30 August 2022 5,337 4 View
Hi, I am using the Masshunter software to quantify my GC-MS data. So, did the integration using MassHunter. but the data (area) are weird. I have a lot of peaks in one chromatogram, therefore,...
10 November 2020 2,576 5 View
Hi, I am planning to do in- vitro assays for production of triterpenoids in yeast. Looking for the proper strain, I found the strain GIL77 suited for the purpose. However, It's difficult to find...
21 August 2020 395 1 View
Dear All, I have SNPs file associated with the sequence of my genes of interest. As the objective is to apply the selection test on SNPs, I need to extract the coding sequences from them;...
26 May 2020 762 0 View
I have downloaded the SNPs from a public database and extracted the SNPs associated with my gene of interest. The output are in several VCF files based on the # chromosome/ scaffold. So,...
17 May 2020 5,498 7 View
I was wondering if we could skip one step in cloning process from sequencing vector to expression vector? Meaning that start cloning the gene directly into expression vector such as pET 28b and do...
05 February 2020 9,960 2 View
I am cloning terpene synthase genes (CDS) there are different types of them in the same plant which can be quite similar too (i.e in some cases about 97% identity). I was wondering if I it would...
31 January 2020 2,356 0 View
I plan to clone some terpene synthase genes into a sequencing vector and later do the heterologous expression using an expression vector. I was wondering if anyone has experience with free...
29 January 2020 7,994 3 View
I am trying to do evolutionary analysis of around 125 CDS alignments using PAML software; it kills the analysis just after few seconds of running. I have used different software to make".phy"...
16 December 2019 2,511 1 View
I am using Rstudio to create heatmap for some expression data; My file includes 38 genes, therefore, 38 rows and 20 columns as samples. The output heatmap shows only 19 of my genes!!! Any idea how...
11 December 2019 7,728 3 View
I need to convert my fasta format alignment file to "Nexus.nxs" to be able to use it in Mybeys. What is your suggestion? So far the files I created using Mega x as well as an online database...
10 October 2019 4,631 2 View
I am running a set of data (DNA coding seqs, alignments done in mega x) in Nexus format. I used mrbayes template (by Barry G hall, Utility program in Mr bayes) to build my block. However, It...
09 October 2019 4,655 6 View
Is anyone can help in how to install trimAL on windows 10? I am gonna use the software for trimming of my aligned sequences. It seems the software does not get installed in my system! Any idea...
16 September 2019 6,324 3 View
Why we add Cytochrome C solution for staining the viruses to be visualized using TEM? I follow this protocol: •VLPs are prepared at concentration of 300 ng/ mL •The grids are washed with...
24 March 2019 2,057 4 View
I am fusing small peptides around 10 to 30 a.a into the C-terminus of a larger protein. The question is if I need to design specific antibodies against those small epitopes? I actually have two...
11 May 2018 1,276 1 View
Hi all, I am looking for some papers/ publications where a Hepatitis A vaccine has been tried or made in any plant. To my knowledge, There are not that much study on plant based vaccine against...
18 February 2018 8,677 1 View
Hi all I am using pET29C for cloning purposes. Considering the fact that it already has an C-terminal His tag, what happens if I include a 6x his in my reverse primer too? is it depending on the...
31 January 2018 8,032 5 View
I have cloned my gene of interest into pET29c, and sequencing verified it too. I used a c-terminus his-tag in my insert. However, when i do induction (1mM IPTG, Over night at 28C), I see a band...
12 January 2018 2,707 8 View
Dear researchers, I have coloned my gene of intetest in pET29 and confirmed it by digestion and pcr from the miniprep. Interestingly, sequencing failed when i use my primers. However, it reads...
18 December 2017 7,359 5 View
Hi all, I am using pET29 expression vector,and use NdeI and KpnI restriction sites as well as XbaI for my fusion protein. The question is if I can use the his-tag of the vector for further...
05 August 2017 1,622 5 View
I am using pET29 for cloning. Digesting with kpnI and NdeI, the result was very good looking on the gel; i did gel purification but i dont see a trace of band after purification! Since i nèed to...
24 July 2017 7,664 4 View
I am inserting a sequence to pET29 and after this ligation I will need to fuse my gene of interest to the same vector. I was wondering if I need to transform e.coli ( strain Bl21) and produce...
10 July 2017 8,326 6 View
I doube digested pET29C with Ndel and Kpnl-HF from NEB. However, after gel purificartion, runing the agarose gel gave me just a single band. I expected to see three bands represnting the isolate...
10 July 2017 8,118 2 View
I am using NdeI and Kpnl-HF for digesting pET29. The digest product will be gel purified and will be used for ligation. I was wondering if i need to heat inactivate the enzyme after digestion? For...
08 July 2017 6,000 9 View
I need to design primers for cloning a gene into pCAMBIA vector. The plan is that the first AUG of the gene of interest must be at +1 of CaMV35 S promoter. I am thinking if any restriction site...
28 June 2017 9,139 6 View
Dear researchers, Expressing my protein of interest in E.coli,I want to use a centrifugation method to purify the protein; I think that way, based on my protein density I will be able to purify...
26 June 2017 7,897 7 View
My plan is to express my gene of interest within pET29 in E.coli cells. However, the expected protein is around 125kD. I was wondering if it's possible doing that? There is an idea behind using...
19 June 2017 7,888 9 View
I am going to clone my PCR product into an expression vector. the PCR product is a clean single band around 600bp; however, I am out of column in my fisher scientific PCR purification kit. I was...
12 June 2017 769 7 View
Hi, I am applying rapid freezing stress like -5 centigrade degree on suspension cells. However, I know the main stream is that to gradually cool the samples down; now am somehow wondering if it is...
26 October 2015 2,040 4 View
Hi Dear Researchers, I am using Nile Red to visualize the lipid droplets of the cell membrane after applying stress on tobacco cells. I suppose not to see the nucleus but there is a clear...
09 October 2015 4,333 4 View
Hi, I want to extract total protein from tobacco suspension culture and used TCA- acetone method which I think was not well for my analysis as the proteins are not solidified well in the sample...
15 September 2015 8,010 3 View
Hi, To determine polysaccharide composition of the cultured tobacco cell walls, I follow the protocol below. http://www.nature.com/nprot/journal/v7/n9/full/nprot.2012.081.html However, the thing...
03 August 2015 7,663 5 View
I want to transform BY-2 cells with the agrobacterium that carries Gentamycin resistance gene and the construct which is kanamycin resistant. In almost all protocols I am looking, two antibiotic...
27 July 2015 7,643 8 View
Hi, I do need a protocol to isolate the cell wall of tobacco cells growing in suspension culture. The isolated cell walls are gonna be used for polysaccharides determination. Any idea is...
21 July 2015 6,447 3 View
Hi, To visualize intracellular organelles with TEM, I want to know if I need a specific marker and/ or a dye to see the ribosomes? Best,
05 July 2015 2,085 7 View
Hi, In my experiment, I have two factors but one of them just has one level since it's not the main factor I don't want to have more levels. I was wondering that with this situation should I use...
21 June 2015 1,564 8 View
Hi, I want to measure ROS activity in the treated cells.I would prefer a way that is more trustful. For this case, I can measure concentration of different ROS produced in the...
13 June 2015 8,947 5 View
Hi, I want to determine the sugar composition and protein content of the cells; as there are different methods to do that, I would be grateful if you suggest your preference methods according to...
06 May 2015 9,031 3 View
Hi, I am using DAPI for screening the nucleus in a tobacco cell line. However, when looking at the cells under microscope, DAPI is visualized mostly in the membrane and somehow in cytoplasm other...
24 April 2015 750 5 View
Hi, I need to know the exact osmatic pressure that different percentages of PEG6000 apply. please share with me any paper related. I think there must be sth like a table. thanks,
17 April 2015 8,877 3 View
I would like to use a fluorescent dye for analyzing tobacco cell nucleus. Do you think which dye works better? best
01 April 2015 2,367 4 View
I need a protocol for DAB staining in the cells. So, any method/ protocol suggested would be appreciated. Thanks
30 March 2015 4,520 3 View
I'm planning to measure reactive oxygen species produced in tobacco cell lines under stress. Which method do you think is better? I see some articles measure ROI; it looks similar but I don't know...
30 March 2015 3,238 2 View
This is a matter of question for me that if we want to screen the cell response to a treatment in a short time, what cells are suitable? I am talking about plant cell lines. However, it might be...
05 March 2015 7,954 3 View
I am going to apply drought stress on the cell suspension culture, using by PEG8000. So, I will add different concentrations of the PEG into the media and then plant the cells in it to see the...
04 March 2015 6,871 8 View
I need some information about corn (zea mays) cell lines. How do they grow? Are they fast growing cells? How often they need sub-culturing, etc?
17 February 2015 6,858 7 View
I treated the cells , BY2 cells, with an organic material but they grow like the control while, I expect the better grow and, this problem is driving me crazy as I'm doing it for about three...
12 February 2015 8,407 3 View
For the cell culture, I am using tissue culture plates; I just noticed that the ones we received recently have this label'' Non tissue culture treated plate''. Is there sth related to evaporation?...
15 January 2015 684 14 View
The BY-2 cells are non green cells and even when they are under continuous light, don't change their color. Can changing hormone in their media can make a difference? I worked with other cell...
01 January 2015 5,723 13 View
I dissolved PEG 8000 in water (670mg/ml). I need to filter it for further use in the culture, but it's kind of gelatin; seems impossible to filter. Can I autoclave it? What concentration you...
28 December 2014 7,758 9 View
Since my cells are growing in well plates and I have only a limited amount of the cultures, let say100/1000 microliter there, it seem the conventional method of dry and fresh measurement is not...
26 December 2014 9,654 2 View
I need to measure cell viability in 48/24 well plates. However, since the cells are under treatment of an organic material which has some absorbance itself, I can't use/ trust the optical density...
23 December 2014 6,829 4 View
I am not quite sure that if I don't deactivate trypsin what is going to happen to the cell suspenssion. Apart from serum which is being suggested for deactivation, what else could be used for this...
02 December 2014 2,937 4 View
I'm studying tobacco BY-2 cell lines, and now i need to find some cell lines like them for zea mays; do yuo think they are available. If not, what do you suggest something like prepared...
05 October 2014 4,277 3 View
