In taxonomy bar plot, I have some Archaea domains while I used bacterial 16S rRNA primer sets. How possible to justify it?
Many 16S primers will amplify some of Archea taxa, but are not usually as inclusive across the Archea as they are across bacteria (see https://mbio.asm.org/content/mbio/8/6/e00824-17.full.pdf )
Hi, In the fiollowing paper I would like to know what is the meaning of mix in dsrB forward primer? Diversity analysis of sulfite- and sulfate-reducing microorg... DSR1762F2 DSR1728FmixB dsrB...
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I have two treatment groups of sample plus a group control. Each treatment and control have own three replicates. I am using Qiime2 for data analysis. I am in dilemma to the following ways below....
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Hello Dear Friends, Hope you are doing well! I have a question regarding the two phrases: 1. reference sequence 2. reference taxonomy What is the difference between them? What makes them...
01 January 1970 2,899 1 View
Could you please tell me for what reasons we should use a cloned DNA segement as a standard to find out how much a certain gene expressed? If you need an example, take a look the Quantitative PCR...
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I need to creat a standard curve for a qPCR assay by plasmid, after plasmid extraction, I run electrophorasis assay to check its quality. I visualized small nucleic acids in the lower layers...
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Dear researchers, As you know, it might be some ambiguity nucleotides in primer oligos appeared. For example, in my case, which I took the forward and reverse primers from a paper, there are some...
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I could not find the PCR amplicon length by a primer set mentioned in a paper. I googled and followed the references, but not worked. Have you got any suggestion?
01 January 1970 7,791 5 View
I was trying to find alpha, beta and gamma diversities in the Brock biology of microorganisms book, but there is no such info in. Similarly, I could not find evenness and richness phrases. I have...
01 January 1970 1,979 8 View
I was asked to provide the 16S rRNA gene primer's name. I designed the primer for 16S rRNA gene with these details. 926F and 1392R primer Pair I googled its name but I did not catch anything....
01 January 1970 8,249 4 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...
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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...
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How to design VN primer to attach with universal reverse primer
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It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...
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Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...
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Hello all, I have been trying to follow a 2-stage PCR protocol used to amplify barcodes of a large yeast library, as per Nyugen et al. (2022) -...
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I want to introduce 2 mutations using Agilent's multi-site mutagenesis kit, I designed the primers using their online tool and it gave me 2 sets for primers (1 set for each mutation) so I was...
25 July 2024 6,517 3 View
Hi everyone, I have extracted DNA from a biogas bioreactor using Qiagen kit and prep cDNA library then used this library as template to optimize primers for qPCR (taken from papers). Some of the...
23 July 2024 1,329 5 View
Hello researchers, Sorry for my stupid question. I am learning the QIIME2 workflow for analyzing some 16s amplicon NGS fastq data. I found a very nice paper with data and code public available...
20 July 2024 5,405 2 View
We are in need of primers for a PCR protocol for chicken genome. We are looking at the COI barcode, 648-bp fragment of the 5' end of COI. Specifically Primers FalcFA, BirdR1, CO1_ExtF,...
16 July 2024 118 2 View