This column extracts DNA and total RNA (mRNA +miRNA), so had a query that whether miRNA so obtained can be used for miRNA profiling.
The protein of my interest is of 69 k Da, but i observe a nonspecific band at 55 Kda along with the specific band, even after increasing blocking i see this band, it also stains ladder at same...
08 September 2014 9,608 4 View
I want to down-regulate gene of interest in SCC cells but this cells are hard to transfect. I have tried using Xtreme gene as well but got 10-15% transfection.
07 August 2014 5,466 3 View
sh-RNA for a gene is cloned in pLKO1.puro vector and it foams a strong hairpin loop, due to which sequencing reaction terminates abruptly. Please suggest me what can be done in this situation.
01 February 2014 5,446 2 View
I am standardizing IHC for pEGFR detection using vector ABC kit, and tried different retrial conditions and antibody concentrations, but I'm unable to get staining. Can anyone suggest any specific...
06 July 2013 215 2 View
I use Nested PCR with MY09/11 followed by GP5+/6+ primers, but if anyone has a better set than these please let me know as prevalence of HPV in gingivobuccal cancer is very low, so I don't want to...
02 March 2013 9,577 5 View
I use PCR to detect presence of HPV in my tissue samples, In positive sample i get single specific band (150 bp) but those samples which are negative i get higher mol wt non specific multiple...
10 November 2012 5,988 29 View
In my study on human tumors, samples showed down regulation of one of my genes of interest. I am interested to know whether its because of promoter methylation or transcriptional inhibition. How...
09 October 2012 2,957 9 View
Normally I use QIAGEN kit for extractions from tumor tissues, but now I am planning to do it from premalignant lesions where I need only epithelial cells. I tried taking serial cryo sections and...
09 October 2012 8,237 21 View
I am planning to block methylation in my culture and check its effect on expression of particular gene, so how should i treat my culture with aza cytidine? i.e should I give continous dose or at...
09 October 2012 1,518 18 View
Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...
11 August 2024 7,082 3 View
I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...
10 August 2024 8,539 5 View
I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...
08 August 2024 9,882 2 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...
06 August 2024 807 3 View
I am planning to collect human fecal samples for metatranscriptomic analysis using MGI. These samples are from indigenous people living in a region with high temperatures. I will have access to a...
06 August 2024 1,367 3 View
How to design VN primer to attach with universal reverse primer
05 August 2024 2,116 3 View
Are there any statistical methods to justify your sampling technique using SPSS or AMOS?
05 August 2024 9,153 4 View
I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...
05 August 2024 6,787 4 View
I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....
05 August 2024 2,570 3 View