Can the PCR fail to amplify high loads of target?

Hello, I did the PCR (long-range, target ~5 Kb) that is shown in the attached file. The lanes are: 1. ladder 2. target (plasmid purified from E. coli after transfection), 50 ng/uL corresponding to...

01 February 2020 3,375 3 View

Is it possible to get the list of bacteriophages against a given species of bacteria?

Hello, from this website (http://phage.org/names/2000/) it is possible to determine the genera targeted by a family of bacteriophages, for instance the phage species DS96 , H24 , M35 , P3 , P9 ,...

06 July 2019 6,924 7 View

How to calculate the phase of a split CDS?

I am trying generate a custom general feature format (GFF) file but I need to calculate the phase of the Coding Sequence (CDS) for split ORFs, that is ORFs that have multiple CDS; for instance I...

09 October 2018 2,753 0 View

BWA-MEM memory use is really 5 GB?

07 August 2018 6,581 0 View

Where can I publish code?

Hello, is there a journal that publishes computer code for bioinformatics purposes but it is not so much high end (no Oxford or Nature class to be clear). The code I have written is not mind...

02 March 2018 3,695 0 View

Set up of Ligation mediated PCR (LM-PCR)

Dear all, I am working at using ligation-mediated PCR (LM-PCR) to amplify DNA of unknown origin; at the moment I am testing the system using amplicons, which should have the terminal adenine at...

04 May 2017 8,592 3 View

What journals in the medical/microbiology area allow to publish without asking fees but also fulfil the open-access requirements of the universities?

10 November 2016 8,485 3 View

Are the any works on the structure of mRNA obtained by NMR or X-ray?

06 July 2016 9,375 2 View

What is the actual size of a DNA nucleotide?

06 July 2016 9,815 0 View

Is there a qPCR quality control reference paper?

It is common knowledge that the accepted range for the efficiencies of a qPCR is between 90 and 110 percent whereas the Y-intercept should be between 33 and 37 cycles (data coming from Life tech...

11 December 2015 9,595 4 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

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

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

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View