What is causing these Taqman probes with the same sequence but different dyes to perform differently?

We are having a perplexing issue with some IDT double-quenched probes. We have a particular amplicon with a SNP and have control DNA that is wild-type, heterozygous, and homozygous for the SNP....

31 December 2015 7,024 3 View

How can I design probes in a GC poor region with a high enough Tm?

05 June 2015 2,884 5 View

What are the alternate loci?

I am currently designing a SNP genotyping assay for CYP2D6 and I am confused about the alternate loci that keep appearing in my blast searches. What is an alternate locus? Does every gene have an...

02 March 2015 1,712 3 View

What does the allele CA|CA in the 1000 Genomes Browser mean?

31 December 2014 1,412 1 View

If sanger results include nucleotides past my reverse primer does that imply I am amplifying unwanted regions?

I sequenced with both the forward and reverse primers in separate reactions (one tube forward primer + amplified DNA, and the other tube reverse primer + amplified DNA). The reverse reaction shows...

11 December 2014 2,838 1 View

Why does PCR with SYBR Green have a high fluorescence at cycle zero?

PCR was run with SYBR Green Taq and three different template DNAs that are wild type, heterogeneous, and homolozygous for a SNP. The only difference between the tubes is the template DNA. The...

11 December 2014 8,964 2 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

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

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

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 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

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