Is the melting curve generated by fluorescent loop-mediated isothermal amplification informative?

19 February 2023 0 8K Report

I recently tested some samples using the fluorescent ring-mediated isothermal amplification method, but the melting temperature of the sample was about 2°C lower than the positive control. Is this due to non-specific amplification of the sample?

Badges
Science topic

Similar topics
Filing

More Fuyou Che's questions See All

Why my plant protein cannot resolubilize after TCA/Acetone extraction?

I am extracting proteins from plant samples and am currently attempting to use the TCA/Acetone extraction method. I am having trouble resolubilizing my sample after drying the pellet. I collected...

15 July 2024 9,025 3 View

Hello everyone, please has anyone expressed proteins using the HEK293 cell line before ?

I tried several times using the Lipofectamine 2000 reagent, extracted the proteins but couldn't detect on Western blot. My plasmids were constructed with PcDNA 3.1+ Please does anyone have any...

20 March 2024 4,533 4 View

If it is possible to use 75% ethanol to measure porosity instead of using 95% or 99%?

Hello everyone, I'm trying to measure the porosity of keratin/alginate hydrogel. I found high value (around 105%-110%) when I try to measure it using 95% ethanol. I have ever tried to use water to...

31 May 2023 3,268 1 View

Speccial Issue: "Soft Magnetic Materials and Their Application". any submission?

A special issue of Materials (ISSN 1996-1944).

19 March 2023 1,490 0 View

Speccial Issue: "Soft Magnetic Materials and Their Application". any submittion?

A special issue of Materials (ISSN 1996-1944). Deadline for manuscript submissions: 20 April 2023

19 March 2023 7,822 0 View

How to verify the specificity of a primer pair?

I recently designed a pair of primers capable of amplifying 12 Chlamydia species with a target fragment length of 184bp, which was confirmed to be Chlamydia-specific by NCBI primer BLAST. I used...

16 March 2023 9,076 5 View

How can I purify monoclonal antibody from cell culture supernatant?

I have obtained monoclonal cells of hybridoma by cell fusion assay, it grows well in hybridoma medium which is GIBCO brand and supplemented with 20% fetal bovine serum, L-glutamine, how can I...

27 February 2023 2,196 3 View

What is ideal range for MSE value for ANN?

The MSE value from my ANN training is over a hundred for water quality forecasting. Is that mean that my ANN model is bad because the larger MSE means the larger the error. for example, the...

21 January 2023 5,273 1 View

What is the reason why the control wells without the encapsulated antigen are positive in the indirect Elisa assay?

When doing the indirect ELSIA experiment, I only added the antigen dilution to the wells without adding the corresponding antigen, but the results showed a high OD, the primary antibody was the...

26 September 2022 3,715 2 View

How to calculate the conversion factor for FFQ if it comes with range?

Hi. For FFQ questionnaire, how to calculate the conversion factor [𝐹𝑟𝑒𝑞𝑢𝑒𝑛𝑐𝑦 𝑜𝑓 𝑓𝑜𝑜𝑑 𝑖𝑡𝑒𝑚𝑠 𝑖𝑛𝑡𝑎𝑘𝑒/𝑑𝑎𝑦] if the option has range? If once a week, the conversion factor will be 1/7=0.14. How about...

20 June 2022 5,761 2 View

Can formaldehyd-fixed gelatin be melt?

I am puzzled about the properties of gelatin, when used for tissue-embedding. Hard gelatin can be melt again by temperatures about 40°C. Formalin-fixed gelatin is like crosslinked protein and...

07 August 2024 1,684 1 View

In the presence of VERY high concentration (1000 ppm) of silicon, the adsorption isotherms of arsenic are showing anomalous trends, why so?

I conducted an adsorption experiment of arsenic on soil in the presence of different doses of silicon as competing ions to see the effect of silicon on arsenic adsorption and desorption. I took 5...

03 August 2024 6,498 3 View

Melt curve problem in qPCR?

Do these peaks in the curve indicate primer dimer or non-specific products?

01 August 2024 7,465 0 View

What is the best protocol for freezing the rat brain in preparation for protein detection by Elisa ?

What is the best way to freeze the rat brain - liquid nitrogen, Dry ice , isopentane - after isolation for later protein investigation by Elisa?

01 August 2024 1,722 1 View

How to Post-fix Mice pup's brains?

I need to Postfix mice brains. I slice the brains using microtome at 40 microns. These are Postnatal days 7, 11, 14, and 21. I am using half the brain for other analysis, so I need to fresh freeze...

31 July 2024 1,121 4 View

Determine pore diameter by BJH of TiO2?

Dear all, l need to help me to determine pore diameter. I got from BJH data the rp(peak area), Desorption Isotherm was 1.6 nm, but from Isotherm l got type lV with hysteresis H2 loop. Can l used...

10 July 2024 2,871 1 View

How to Freeze Embryos for Spatial Transcriptomics?

Hello everyone, I am currently working on a project involving spatial transcriptomics on E14 mouse embryos. We need to perform cryosectioning without fixing the embryos. Here is the protocol we...

10 July 2024 5,984 0 View

Suggestions for storage of CD4+ naive splenocytes for future use?

Hi! I am working on a biophotonics project in which I need isolated but inactive CD4+ splenocytes. Every protocol and piece of literature I have found immediately activates and cultures...

02 July 2024 4,887 0 View

How does the melting point of a spherulite correlate with its size?

For Polybutylene Terephthalate (PBT) it is known that if you cool it down at fast rates to have an additional melting Peak in DSC-Scans. Androsch et al. (2015) conclude that this is due to...

30 June 2024 2,891 1 View

Why does a nitrogen adsorption-desorption isotherm form two loops?

BET analysis

27 June 2024 6,964 1 View