Would infusion clontech be the best approach for a sequence 4k long for 5race and 3race and cloning?

More Louis Tong's questions See All

Is species of the antigens important for immunisation?

oes anyone know if I want to immunize Lewis rats with lacrimal antigens, whether it is important to use lacrimal extract from a different species like SD rats, or even from C57B6 mice?

31 December 2015 3,795 3 View

Question on animal models of disease?

Does anyone have experience doing a delayed hypersensitivity type model in the eye?

31 December 2015 7,682 1 View

What is a good ELISA kit for histamine, tryptase or chymase?

Need to test on tears (small fluid sample)

31 December 2015 5,588 1 View

Is methylene blue staining specific for fibroblast colonies?

Thanks in advance.

02 March 2014 4,218 2 View

Can anyone give some guidance on DNA sequencing, reaction and services?

If I have the reverse transcription products from total cellular RNA, is it possible to sequence fragments with specific primers without gel purification of the cdna first?

09 October 2013 5,015 5 View

Does any one know the best place to get hydron pellets for corneal pocket model of angiogenesis?

Seeking advise on animal model

07 August 2013 5,478 0 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

No title

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

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View