What is your favorite polymerase?

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Does anyone have a preferred spectrophotometric detection method for arsenite vs. arsenate?

So far we have tried a variamine blue assay from Narayana et al. 2006 and a silver nitrate precipitation from Simeonova et al. 2004. In the variamine blue assay instead of intensity correlating...

05 June 2016 2,690 2 View

Does anyone have B. subtilis that is constitutively expressing a fluorophore?

I am looking to co-culture/compete some B. subtilis with other microbes. I am new to this model organism and the field so I don't yet have any contacts to ask for strains. Does anyone on this...

07 August 2013 1,193 3 View

What is your favorite scientific paper of all time?

I am helping to teach a course on communicating science and through this course, I have come into contact with an absolutely beautiful paper. It is clear, concise, and elegant. I am not at all...

09 October 2012 2,715 3 View

How do you feel about genetically modified organisms? What do you think you know about them?

I was at a scientific conference this weekend and was surprised to hear about the impact that anti-genetically modified organism (GMO) feeling has had on funding and direction in the field of...

08 September 2012 1,453 14 View

Is anyone familiar with SimBiology (MatLab) and can you describe a functional conditional statement?

I am a biologist who is trying to become more quantitative as well as more model savvy. One excellent tool I have stumbled upon is the SimBiology toolbox for MatLab where you can graphically...

08 September 2012 5,432 3 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 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

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 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

I want to performe MALDI-TOF MS for bacterial identification, I would appreciate it if anyone can tell me the matrix solution preparation protocol ?

based on what I've studied on multiple papers, for peptide and protein identification Alpha-cyano-4-hydroxycinnamic acid (CHCA) is an appropriate matrix to use, I also know that the CHCA powder...

26 February 2021 6,799 3 View

Jian-bo Wang

Primeratar

Andres Esteban Marcoleta

Hello, which polymerase you should use depends on what you want to amplify. If you need accuracy the Pfu series works fine, If you want to amplify large fragments (i.e. 5 kb or more) you may use a mix of Taq and Pfu polymerases. If you want to amplify standard-size fragments and accuracy is not so relevant, the best choice is Taq. Regarding brands, we use Taq Polymerase recombinant (Invitrogen) and Pfu ultra II Fusion (Agilent).

Regards

Yu Xia

We usually use enzymes from Takara。If we care about accuracy and efficiency, the KOD enzyme from Toyobo is preferred.