I get Non-specific products & multi-peak Melt Curves with my Real-Time PCR procedures, any advice on solving this problem?

I still get Non-specific products & multi-peak Melt Curves despite changing the annealing temperature of my designed primers several times (58C, 59C, 60C, 61C, 62C, 63.7C & 65C). I've also...

11 December 2019 3,112 6 View

Can i run a single RealTime PCR experiment with 2 different Annealing Temperatures?

I have 2 different pairs of Primers, the annealing temperature of the 'reference pair' is 54C, while the temperature of the other 'test pair' is 60C. I'm running my experiment on the device of...

09 October 2019 2,002 4 View

What's the minimum purity of a genomic DNA sample used for Zymo Research's OneStep qMethyl-PCR Kit?

The purities of my genomic DNA samples range between 1.51 and 1.71 at 260/280nm wavelengths. This is the kit: https://www.zymoresearch.com/products/onestep-qmethyl-pcr-kit

02 March 2019 8,146 3 View

I need an explanation for the following 8 DNA Chromatograms?

- 4 of them have a Homopolymeric Region followed by Noisy Multiple Peaks. An example image for one of the samples of this case is attached with both the sequences of its Forward and Reverse...

06 July 2018 6,834 7 View

Salting Out Method for Salivary DNA?

I use Chelex100 for extraction and MEGAquick-spinTotal for PCR purification. Some samples were very faint so i need to make salting out. Thanks in advance.

08 September 2017 6,164 4 View

PDF Protocol of Bosphore Purification Kit?

07 August 2017 8,554 2 View

Any alternative for BioEdit sequence alignment editor?

07 August 2016 8,423 7 View

What is the suitable temperature for storing saliva for DNA extraction and Vitamins B test?

06 July 2016 3,238 4 View

Is the OneStep Methylation Kit suitable for DNA extracted using Chelex Extraction Kit?

05 June 2016 7,224 0 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

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

Which detection method is better for an ELISA-based assay: colorimetric or fluorometric?

Hello, I am planning to perform an ELISA-based assay to quantify the percentage of genomic methylation (5-methylcytosine) in human embryonic stem cells. I've been searching for some kits and I...

16 February 2021 8,389 3 View