How do I design primers/probes for eukaryotes?

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How do I prepare an RNA gel?

I am looking to assess the quality of some RNA that I have extracted from human adipose tissue. Currently using a page supported by ThermoFisher...

09 October 2018 4,808 0 View

How to design good primers?

Other than assessing for temperature of hairpin structure (for both forward and reverse primers), self-dimer Delta G, and hetero-dimer Delta G, is there anything else to look at before ordering...

09 October 2018 7,711 4 View

Are my adipose tissue samples still viable after being in a -80 that went down on us over the weekend?

I have some human adipose tissue samples. -80 C went down on us over the weekend. We are unsure of how long the freezer was actually down for. When we got it back on the outside thermometer on the...

07 August 2018 6,567 6 View

What is the amount of RNA (adipose tissue origin) used for 1-step RT-qPCR?

I am working with human adipose tissue and want to quantify the expression of a few genes. My intent is to use a 1-step RT-qPCR protocol/kit to do this. I am not able to find information on this...

04 May 2018 5,609 3 View

What is the best way to store protein extracted from adipose tissue samples after QIAzol homogenization?

I will be extracting mRNA, miRNA, and protein from human adipose tissue using the miRNeasy mini kit and the RNeasy mini elute cleanup kit. After phase separation I want to store the proteins...

03 April 2018 4,490 3 View

What is the best kit/reagent to use for extracting total RNA and miRNA from human adipose tissue?

I will be extracting mRNA and miRNA from human adipose tissue. In your experience, what have you found to work the best?

02 March 2018 2,577 4 View

Issue extracting RNA from adipose tissue

I am having issues extracting RNA from adipose tissue. Currently, I am using the miRNeasy Mini Kit from Qiagen following their protocol. The result is a low concentration and an A260/A280 of...

01 January 1970 9,418 2 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

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

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

Can we convert a thousand of FASTA sequence in numeric form in .csv format? If yes kindly send me the script for the same?

I have a .text file for various FASTA sequence , and i want to convert these sequences into a numeric file which will be in .csv format. OR I want to extract physiochemical properties(features)...

25 July 2024 3,650 2 View

Do you mix and add all primers together in a singel PCR reaction (4 total primers for 2 mutations) when using Agilent's multi-site mutagenesis kit?

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

Can I please ask why my samples from anaerobic bioreactor giving me different size PCR product even after multiple runs?

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

Could someone please provide a list of journals that accept the application of methodologies in nature-based solutions?

I am looking for journals that admit the publication of results from applying the IUCN Standard for designing and monitoring Nature-based solutions. Many thanks for considering my request.

23 July 2024 6,444 2 View