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Questions related from Sailesh Palikhe
I am trying to perform HE staining of small intestine. I used OCT solution to make sections. After HE staining when I look my slides under microscope, I do not get intact villi. In much of the...
02 February 2020 5,893 3 View
We are currently working on a cytosolic protein; however, we think that this protein also has a secreted form and can be detected in the bloodstream. As there is no ELISA kit available for this...
02 February 2020 413 2 View
I want to study whether two proteins: Myc-tagged protein A and Flag-tagged protein B, interacts with each other or not. For that I co-expressed these proteins in HEK293T cells, and later (after 48...
02 February 2019 1,884 4 View
I want to determine whether protein A and protein B interacts with each other by co-immunoprecipitation analysis. For that I prepared Myc-tagged protein A and Flag-tagged protein B plasmid DNAs. I...
02 February 2019 944 4 View
I am just wondering whether different labs prepare immortalized bone marrow derived macrophages (BMDM) in their labs. I have experience of preparing primary BMDM from mice femur, but have no idea...
01 January 2019 8,906 2 View
I have been working on two proteins. And I think that these two proteins interact with each other. To prove that, I analyzed there association by endogenous IP, but unfortunately I could not get...
12 December 2018 9,716 4 View
Articles published in journals contains both reference and citation, while books contain only reference. Does anybody have idea about it?
10 October 2018 894 0 View
I just started to do EMSA to check interaction of a protein and DNA. When I checked the binding buffer used for EMSA, I found that EDTA is also used. I know that EDTA is a chelating agent, and...
05 May 2018 2,662 1 View
I have been recently trying to isolate primary microglia from mice pups brain (P0-P1). To prepare mixed microglia culture, I used 3 brains for a 75 cm2 flask. After around 10-12 days, astrocyte...
04 April 2018 3,747 12 View
Currently, I am doing knockdown of certain genes by shRNA technique. We use lenti-virus for this process. The problem is that we are getting low titre. The experimental procedure is: Day 1: Seed 7...
05 May 2017 2,400 6 View
I am culturing an adherent cell in a 6-well plate. Unfortunately, one of the well got contaminated. Now, I need to change the media, and I just afraid while doing so the contamination get into...
09 September 2016 7,868 6 View
Does anybody use 7500 Real time PCR of Applied Biosystems. I want to use touchdown PCR setting but don't know how to set. Can anybody help me?
04 April 2015 3,326 5 View
I want to over express a protein in different cell lines of human. I know that we can use cDNA for it. But, I wonder is it possible to use just ORF (cDNA without UTR regions) for cloning in a...
01 January 2015 1,645 5 View
I want to induce MRP4 in different cell lines by using some specific inducing agents. Does anybody know about any specific inducing agents for MRP4?
01 January 2015 9,479 1 View
I am going to use real time PCR using Taqman assay. I have experience of RT-PCR by using SYBR but I have no idea about Taqman. Can anyone suggest a protocol for Taqman?
12 December 2014 7,498 7 View
I am amplifying 2 kb of a gene. For that i used Taq polymerase enzyme and 22 nucleotide length primer. I checked the amplified sequence by sequencing and it was fine. But when I amplified 2 kb of...
12 December 2014 5,983 8 View
I want to check the effect of an SNP (single nucleotide polymorphisms) on gene expression. For that I amplified two PCR fragments containing two different alleles. And then I ligated into PGL3...
12 December 2014 8,637 2 View
Is it possible to know whether the DNA samples contain homozygous alleles or heterozygous ones?
12 December 2014 7,389 7 View
I want to measure LTC4 release from HMC-1 mast cells. It would be better if someone could share their experience. I want to stimulate mast cell to release LTC4.
11 November 2014 4,968 4 View
I need specific inhibitor of MRP4. I searched many papers but only found non-specific inhibitors. Can anyone suggests me?
11 November 2014 4,739 2 View
There is a SNP in a promoter region of a gene. I want to know whether any transcription factor binds in that particular region (in SNP). There are many databases which can predict transcription...
11 November 2014 4,676 5 View
I am going to check effect of various SNP of different genes on disease susceptibility. For this, I need to use Multifactor Dimensionality Reduction (MDR) software to analyze gene-gene...
09 September 2014 5,098 1 View
Do we need to incubate suspension cell before treatment with drugs (or any stimulants)? As I know for adherent cell, it is required so because it need time to get attached. Does it make any...
08 August 2014 4,602 3 View
I am trying to amplify 333 bp of promoter region of PTGES gene. I got the required band after PCR but intensity is quite low. I tried with the same condition and reagents that I used for...
07 July 2014 9,195 16 View
I have a promoter SNP of ABCC4 gene (-1393T/C). I want to find rs number of this SNP. Does somebody have any idea regarding this question?
06 June 2014 1,397 2 View
Can anyone explain the Fasta and Attonated gene sequences? Are they the same sequence or different?
04 April 2014 4,683 1 View
I am doing cloning now by using infusion cloning method, but I am not getting good primers. For infusion primer, we have to add 15bp homologous to 5'. When I get a specific primer, then it has...
02 February 2014 9,528 3 View
I have a nucleotide sequence of a gene (short fragment) and I want to know which gene this sequence belongs to. Any suggestions?
02 February 2014 8,999 4 View
Usually for genetic association analysis there are lots of SNPs, but we generally select few tagSNPs based on LD value (r2)? How can we calculate the r2 value to know which SNPs are to be chosen?
02 February 2014 5,770 2 View
I was using chipbioinformatics tools for getting nucleotide sequence, but currently it is not working. Can anyone suggest an option?
02 February 2014 5,200 12 View
I want to perform transfection by electroporation method with Hep3B cell line. Has anybody tried this before?
12 December 2013 5,106 4 View
I performed Gradient PCR to find the appropriate annealing temperature, and I got the high concentration band at 58 °C. But later when I performed RT-PCR, I got non specific band, even sometimes...
12 December 2013 2,657 13 View
I need a little help in naming of SNP. What does "1123" indicates in SNP name, such as -1123C/T? Does this mean upstream of starting point (ATG) or from exon 1?
12 December 2013 7,865 10 View
I want to perform transfection in HepG2 cell line by electroporation method. Does anybody know about electroporation condition? How many cells should I use per well in 12 well culture plate?
11 November 2013 6,015 0 View
As I know, there are two naming ways for SNP, one is on the basis of position of SNP on gene, for example +1123C>G and the other is rs2488457. In second naming system what is the meaning of number...
09 September 2013 5,014 2 View
Can anyone tell me why SNP of intron region are selected for genetic association studies? What is the importance of intronic SNP?
09 September 2013 6,709 6 View
