Protocol PCR primers narG1950m & narG2050m?

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Does anyone have a protocol to transient transfect HEK293 with 2 plasmids?

I need to transient transfect HEK293 cells with a plasmid containing a luciferase tag and a plamid containing a histamine receptor. I have never done this before so I am looking for a straight...

08 September 2020 845 3 View

How to apply thermal and structural loading for each time step in thermal-structural analysis in ANSYS APDL?

I'm simulating an experiment in which a wood beam is submitted to thermal and structural loads at the same time. The beam is put in a furnace where tempeature rises according to ISO curve and at...

18 June 2018 8,802 9 View

Does anyone know if pKD46 system works in E. coli BL21 or DH5alpha to KO a gene?

I need to KO a gene in my bacteria and I'm currently use BL21 strain also for the over expressed one. I've been trying many systems to ko my gene but so far I didn't get any good result and I'm...

03 June 2018 2,089 6 View

Non-neurotoxic brain dye for infusion?

Dear all, I'm looking for a non neurotoxic brain dye that I can infuse into the ventricles through osmotic pumps, to be sure that I'm doing the infusion in right way. The mice must survive for at...

29 August 2017 5,509 3 View

Renal sympathetic nerve staining

I know it's commonly done with tyrosine hydroxilase but I was looking for an alternative IHC (less expensive) method. I came to understand silver staining or Luxol staining are possibilities but I...

12 June 2017 6,112 2 View

Which method do I use to find the best gene reference to use un the RT-qPCR?

Hi, I need to find a gene reference to use in the gene expression of cellulolytic genes in Ascomycete. Which gene reference is possible ti use? Or which is the method to find it? Thanks Giorgia

31 May 2017 9,655 1 View

It is possible to express the increase of the expression of a microRNA as the sum of its intracellular content and the one in the supernatant?

I need to quantify the total expression of a miRNA that is contained in a cell and is also released from it in the culture medium. How can I do? Do You know a few articles in the literature that...

12 December 2016 1,073 3 View

RT-qPCR protocol one or two steps?

Is best to prepare the RT-qPCR in one-step or two-step?

26 November 2016 9,555 8 View

I'm looking for an online atlas ofthe human brain that allows input of xyz coordinates in MNI space to determine and label corresponding brain areas.

Can anyone point me in the right direction here please? many thanks

11 September 2013 6,106 11 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

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

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 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

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

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

Zhi Xiong Chong

Other than primer problem, you may also need to consider whether the DNA/RNA extraction protocol has problem, PCR master mix? PCR condition was it optimized? Consider each factor one by one to troubleshoot.

Ahmed E. Dhamad

Giorgia, please check this website. It will tell you whether your primers have high percentage of off-target and other features which may help you to troubleshoot you problem. Good luck

Giorgia Pertile

Thanks so much everybody. I found the problem was in the sequence of the foward. The start of the correct sequence is TAY, the my primer was TAGC. I order the new foward and I will try.