Transfection vector for stem cells silencing?

i'm currently looking for what would be the best vector/plasmid for silencing genes in rat stem cells culture. I found some paper where they use lipofectamine, plus reagent, vector pCU and pCH. I...

01 February 2017 8,692 1 View

Why can't i get ladder resolution?

I attach a photo of my agarose gel.. As you can see the DNA ladder runs like "all toghether" (sorry for bad english). I followed the manufacturer recomendations (1,5% agarose, 90mV, 1 hour)... The...

05 June 2016 4,582 11 View

Why didn't PCR work?

I'm doing RT-PCR from rat catilage tissue, and the thing is i searched for tissue specific markers such as SOX9, Aggrecan and type 2 collagen. this is what it looks like, the last 3 lanes are the...

03 April 2016 7,328 10 View

How can i get rid of bright background on agarose gel??

I'm running my pcr products in an 1% agarose gel made with TAE buffer. The volume i use is 20 ml with 1ul EtBr. I attach a picture. I want to get as dark as posible for the background and as...

01 January 1970 9,009 11 View

BSA-AGE without centricon?

i need to prepare BSA-AGE for my stem cells culture. The problem is i have centricones 100 kDa, i the ones i need are the 10 kDa ones. Is there another protocol for preparing BSA-AGE without...

01 January 1970 9,775 3 View

Looking for Materials with high thermal expansion coefficient and high temperature resistance?

03 March 2021 8,272 1 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 have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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