Melting curves with the same shoulders but with different primers, possible reasons??

I am getting crazy about this. I am having less than perfect melting curves, with different primers. The funny thing is that while primers give a main peak at different temperatures, as it is to...

07 August 2018 9,002 13 View

Thermodynamic Oligo Alignment vs Traditional secondary structure alignment, who is the winner?

This is an option in Primer-Blast to select which model to use to calculate the propensity of the primers to form secondary structure. I am puzzled about which option is better, the output of the...

10 November 2017 556 0 View

Long term storage and handling of RNA, what about 66 to 75% EtOH without Na Acetate?

HI, I know that RNA is ideally preserved long term after precipitation in EtOH and Sodium Acetate, and it is successively washed in 75% EtOH to remove salts before rehidratetion. I have also read...

07 August 2017 1,265 10 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 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

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Errors in DNA sequences?

I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...

24 February 2021 4,397 3 View

QPCR reaction efficiency with odd results what does this indicate?

I am working on a CFX 96 machine to determine reaction efficiency of my gene of interest. After performing a serial dilution of my cDNA template, I was able to find great reaction efficiencies for...

23 February 2021 809 3 View

What might be the reason for obtaining two bands in plant genomic DNA extraction?

I got a thin band and a thick band i guess it would be the genomic DNA and the 260/280 ratio is 3.

21 February 2021 6,523 6 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

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17 February 2021 4,217 3 View