How to design 5´ primer tag?

09 November 2023 0 4K Report

I have to perform an amplicon sequencing analysis (metabarcoding) with the plant primer trnL intron g and h. For the sequencing I need to add 5´ tags to the primers. There is a methodology to add it?

Badges
Science topic

Similar topics
Chemistry
Biochemistry

More Diego Cruz's questions See All

I got 02 moderator values defining Johnson-Neyman significance regions. Is it an error?

Hi everyone, I am conducting research for my Master's thesis. I am using PROCESS by Johnson-Neyman to analyze my Moderator model. I test the relationship between Public Service Motivation and...

03 March 2021 2,350 2 View

How to include original NMR data in a manuscript?

hello. i am junior researcher. I received a comment from a reviewer that he could not find NMR data in my submitted paper. I had included the NMR graph with explanation and formula to calculate...

27 February 2021 734 4 View

What influences inter-day reproducibility in UPLC-MS/MS analysis?

I was analyzing some metabolites in human plasma. While the intraday precision of these metabolites was acceptable ( less than 20%), the interday precision was quite large. So what influences...

13 February 2021 9,183 9 View

Where can I find FDI data by sector/industry for Asian countries like India and China for the last 30 years?

Common databanks like UNCTAD, OECD, WorldBank and ASEAN Website did not have the data for the last 30 years.

09 February 2021 4,701 4 View

Sample Size for Quasi-Experimental Quants using one-tailed t-test?

Hi, sorry I'm stumped. I'm going to do a study on the leadership styles (using the MLQ Form 5X) of minority women administrators in medical schools. How will I calculate the sample size? Thank...

30 January 2021 2,865 4 View

Which statistical test to use for comparing percentages between multiple groups?

Hello everyone! I am looking for some statistical help! I am working in in vitro micropropagation and one of the aspects that I am studying is the rooting of plants in 10 different growing media....

14 January 2021 7,575 5 View

How to solve AMPS 1D simulation error?

I am trying to model a photovoltaic device using AMPS-1D software. But once I am submitting it to the queue, it remains there for an infinite amount of time without giving the result. I couldn't...

11 January 2021 9,801 2 View

What are core advantages of 2nd & 3rd generation panel data techniques over the 1st generation techniques?

I am interested to know about the difference between 1st and 2nd and 3rd generation panel data techniques.....

31 December 2020 7,204 7 View

Do chemical and physical properties influence on the size of a polymer particle?

I would like to know if the properties of a polymer impact on the size of the particle it forms. I'm working with different polyssacharides (alginic acid from different seaweed), which are...

29 December 2020 7,333 6 View

Does anyone have the Wiley Journal word Template?

Hi I need a word template file of Wiley online library because it is not available on their website.

27 December 2020 7,208 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