Efficient primers to do qPCR for MS2 bacteriophage?

More Guillermo Domínguez Huerta's questions See All

When do you want to use full protein sequences or only conserved domains to make a phylogenetic tree?

Hi, I'd like to know what difference makes using FULL PROTEIN SEQUENCES versus PROTEIN CONSERVED DOMAIN SEQUENCES in order to make a phylogenetic tree. In my case, the goal is to put in...

03 April 2019 8,888 5 View

Model creation in GeneMarkS?

Hi all, I am trying to perform self-training with GeneMarkS to improve protein calling from virus genomes and transcripts. Could someone tell me if it is correct what I am doing? First, I...

09 October 2018 10,017 0 View

What is different in aged ferric oxide?

Hi all, I'm looking for the expert opinion from a chemist, since I am only a biologist. I have a set of samples made in the following manner: many litters of seawater were...

07 August 2018 2,948 3 View

RnaSPAdes for RNA viruses?

Hi all, would you say that rnaSPADES is a better assembler to discover RNA viruses from read datasets THAN SPADES? thanks!

06 July 2018 8,682 2 View

Conserved domain database?

Hi all, I looking for RNA viruses from datasets. I already performed a blastx of the contigs generated against the database of all reference RNA virus proteins download from GenBank. Now I'd...

05 June 2018 10,139 3 View

RP-SISPA with RNA virus isolates?

Hi all, I just did my first RP-SISPA. I extracted RNA from two virus species by different methods. One virus genome is ssRNA of 3 kb and the other virus genome is segments of dsRNA (3, 4 and 6...

03 April 2018 5,504 4 View

Low efficiency of qPCR?

Hi all, I've been having trouble increasing the efficiency of a couple of primers using a standard curve. - The standard curve is built with log10 dilutions of plasmid into which the amplicon...

02 March 2018 9,732 7 View

What are the bioinformatic tools for collecting RNA viruses from metagenomic data?

Hi all, I wondering about bioinformatic tools to capture "RNA viruses" from environmental sequence datasets. dsDNA phages are pretty abundant and have relatively conserved hallmark genes....

02 March 2018 7,041 6 View

Precipitation after autoclaving?

Hi all, I know this is a typical phenomenon, but after reading I'm not getting a convincing explanation. I prepared 4 L of a buffer containing 10 mM KH2PO4 and 1 mM MgSO4*7H2O. This time I...

31 December 2017 2,746 4 View

Iron ions mediated nucleic acids degradation?

Hi all, I looking for literature evidence for this phenomenon. Apparently, RNA (I'm not sure if also DNA) is cleavaged in a non-enzymatic manner by iron. I would need some theory to support...

31 December 2017 7,451 3 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

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

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

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

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

Could dyes amplify the spectrum of light to a specific wavelength?

I am interested to know the behavior of dyes toward light. Specifically, Blue dyes re-emit the spectrum, especially from the green zone (known as principal in LED lamps, and blue dyes are known...

05 August 2024 3,290 1 View

What is the problem with these tissue culture plants?

All plants are green but some of these plants becomes yellow. I did not found any reason. Please help me to find out the real problem.

01 August 2024 589 4 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

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,...

01 August 2024 4,673 4 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

PCR showing no bands - Master mix and primers didn't mix?

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...

31 July 2024 2,406 6 View

How to retain amplicon yield after Ampure bead cleanup, following a 2-stage PCR protocol?

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) -...

30 July 2024 841 2 View

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View