Can i use the miRNA Sequence (given in miRBase) as the forward primer or design the stem loop primers?
I am using Agilent miRNA first strand synthesis kit (cat#600036) for preparing mi cDNA from total RNA. A universal primer is provided with the kit sequence of which is not given. i want to study...
07 August 2018 2,279 2 View
Hello everyone ! I am using mirVana miRNA isolation kit for the isolation of total RNA containing small RNA. Further i used Agilent first strand synthesis kit(cat#600036) for the synthesis of...
07 August 2018 800 5 View
i tried gradient PCR for a new set of primers and got the amplification. However, when i tried running the same set of primers on a single annealing temperature, it showed no or weak...
10 November 2016 9,289 14 View
I am trying to amplify my genes of interest.Though I got the amplified product of the desired size on my first attempt , i am not able to get the amplification again. What might be the problem and...
07 August 2016 9,484 3 View
I have been using mirVana miRNA isolation kit for total rna isolation and had a good experience using that. Recently, i am facing problem with the kit. 280/230 ratio and concentrations are too low...
03 April 2016 5,063 3 View
I am trying to isolate RNA from Breast tumor and adjacent normal tissues (stored in RNAlater at -20 C) using Trizol method. After adding isopropanol , I am getting a transparent droplet like...
02 March 2016 1,479 3 View
I am performing real time pcr for ACTB. I am getting a ct value of 17-20 in my samples. However , i am getting a peak in NTC too with a ct value around 33-35. Also the dissociation curve for NTC...
10 November 2015 3,796 8 View
I am getting two bands below 100 bp in addition to the gapdh amplicon . What might be the reason for these bands? Lane 2- positive control lane 3-6 - samples lane 8- negative control
06 July 2015 4,145 6 View
I am trying to predict miRNAs against the gene of interest using online tools like Targetscan, miRanda ,miRsystem and DIANA micro T- CDS. As these tools produce a long list of miRNAs that might...
06 July 2015 8,395 6 View
I have to perform qPCR for miRNA isolated from colorectal tumor and adjacent normal tissue samples. Is U6 snRNA a good endogenous control for qPCR?
05 June 2015 3,690 9 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
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 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
Hello, Can anyone provide me with the absorption coefficient of methane gas at 7.7 um? Any reference?
06 August 2024 980 5 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
Please can anyone support with the survey questions based on RQ measures and propose how to do it in FMCG industry and include as well the role of brand equity Thanks
06 August 2024 949 0 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View