Have anyone tried SYBR green with GoTaq DNA Polymerase for a qRT-PCR?

Hi, everyone. I need to perform a real time reverse-transcription PCR in one step. What we have is SYBR green I PCR Master mix with GoTaq DNA Polymerase. So far, our attemps to perform the assay...

07 January 2021 6,507 3 View

Does international survey based researches needs approval in each country ethics commitee?

We plan a contingent valuation of ecosystem services. The study will be based on surveys. We want to apply it in fifteen different countries. Is it necessary to approve the project in ethics...

25 June 2020 8,099 3 View

How to choose the best smooth algorithm for your data in Origin?

I have several curves of applied potential versus electric current with hundreds of points in each of them. I am trying to analyze their first and second derivatives, but they are too noisy to...

27 April 2020 5,932 2 View

I can't make my RAPD-PCR work anymore. Can anyone help me?

Guys, I REALLY NEED HELP At the end of the last year I optimized every parameter of my RAPD-PCR, and as a result I did obtain great products multiple times. Then, I went home for my vacation (I...

18 February 2020 9,236 3 View

Hi, I would like to solubilize a hidrophobic compound in order to administer it to mices. Should anyone suggest a nontoxic vehicle?

The compound is very insoluble in aqueous solution and the compound administration will be given orally. The compound manufacturer has no information regarding the solubility of the compound....

16 January 2019 834 1 View

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Hello everyone! I have been consulting this webpage http://beast.community/ and http://www.beast2.org/ , but the examples for the use of BEAUti and BEAST are for old versions. The versions of...

02 April 2018 3,942 3 View

Two-step cluster in SPSS - how to validate results?

Hi everybody. After applying a two-step cluster in SPSS, involving both continuous and nominal variables, how can I validate if the results are satisfactory? In other clustering methods there are...

03 October 2017 5,765 8 View

Have you COI sequences of Potamotrygon orbigyi?

24 August 2017 6,634 2 View

How many sequences it needed to put in a crown group to calibrate a tree?

17 August 2017 4,155 2 View

How can I find out if variable A moderates the effect between variable B and C using SPSS?

I already proved a negative relationship between variable B and C, however my hypothesis in my research paper is that variable A moderates the negative effect of variable B on C. I read a number...

06 December 2016 2,698 4 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 View

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

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 calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 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

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

Genescan gives 3 peaks instead of one?

Hi all, I am doing a genescan analysis for a deleted exon in the patient sample. The amplified region is 215 bp. In the patient sample because of homozygous deletion, no peak is observed while in...

23 February 2021 5,645 1 View