PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

Thin cut brain tissue flaking off of slides?

Hello! I have just encountered a new issue with some tissue flaking off of my slides and wondered if anyone else has experienced this or has any suggestions? The sections were cut at 18 microns...

09 December 2020 4,954 2 View

Why does a Nested PCR protocol amplify samples and not amplify positive control?

I have this problem. Today I used a new positive control of an isolated NC-1 strain in cell culture and the problem persists. I performed a serial dilution test to test the sensitivity of the...

08 December 2020 6,252 2 View

How to avoid the formation of traces and dimers in DNA gel electrophoresis?

07 December 2020 3,052 3 View

Is the pipetting order of the positive and negative controls in the PCR important to avoid in a possible contamination?

I have been watching bands form on my negative controls. I have already changed all reagents, optimized the hygiene of materials and the environment and increased care in pipetting.

07 December 2020 9,090 3 View

Best methodology to calculate food footprint in a university campus through students inquiring?

Hello, Considering sustainability and environment negative impacts of animal based products, I would like to calculate food footprint in a Portuguese university campus through students...

05 December 2020 9,098 3 View

How to standardize and obtain a good PCR reaction with samples with a low and variable amount of DNA?

I work with the detection of N. caninum DNA in milk samples in milk samples. The DNA concentrations in my samples range between 2-90 ng / ul. I would like to establish and standardize a final...

01 December 2020 8,792 3 View

Sample size in G mellonella studies?

Hi everybody, I am working with G mellonella, and I know that one of the advantages of it is the lack of ethics commit and the 3R principles. However, I am very insecure about the sample size I...

29 October 2020 2,079 1 View

COVID-19, Human behaviour and wildlife impacts?

COVID-19 is causing a radical changes in Human behaviour around the world. Mandatory or voluntary isolation for months, suspension of the active management, etc.These changes will have positive...

24 March 2020 4,039 32 View

There are any component of gram + bacteria cell wall that can be used as a positive control?

Hello, I am going to evaluate the immunomodulatory effect of a gram positive probiotic bacteria in vitro and I need to use a positive control. I was thinking about LPS, but it's a component of...

11 March 2020 7,369 3 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View