What's the longest insert you put into a plasmid via quickchange-like methods?

We would like to put a 12-20 aminoacid linker into a recombinant protein expressed in E coli. We've had good experience with Quickchange-like protocols; but never tried with 36-60 nucleotide...

02 March 2019 3,185 4 View

Is Lithium Lauryl Sulfate OK to run K+-containing samples on SDS-PAGE?

Hi all, Can one use Li Lauril Sulfate (LDS) in loading buffer to run protein samples containing high concentrations of KCl, on SDS-PAGE? Does one need to replace SDS in the running buffer and in...

01 February 2019 1,614 6 View

Can you recommend a tabletop front-loading autoclave?

Can anyone recommend a front-loading tabletop autoclave which could be used to sterilize up to six 2L flasks at once with steam? Looking for something that is robust, simple, and ages well (i.e....

07 August 2018 6,653 0 View

Is there a chemically defined, FBS-free media formulation for HCT-116 and U2OS cells (adherent human cells)?

FBS-free medium is what I am looking for. If defined growth factors are required, I can express them (if they are available in recombinant form). Artificial meat companies seem to have figured it...

06 July 2018 7,101 4 View

Suggest a method for harvesting of adherent cells from hollow fiber modules? Does scraping work? Would using hydrophobic fibers help?

We are looking for a cost-effective way to grow adherent cells (HCT116, US2OS), in amounts of 1*10^10, for subsequent isolation of nuclei and preparation nuclear extracts per Dignam 1980....

05 June 2018 7,168 5 View

Has anyone tried using standard hollow fiber PES hemo-dialysis cartridges to culture adherent mammalian cells?

The surface area (1-2 sq meters) and the basic specs (excluded volume, PES memberane chemistry, pressure drop, non-fowling, sterile, QC, Mw rejection) of hollow fiber dialysis filters appears...

05 June 2018 9,437 0 View

Can 5+M guanidine-HCl denature dsDNA PCR products upon prolonged exposure (>16 hours, room temp, this is for crush and soak of acrlamide gels)?

I am wondering if prolonged incubations in 5M GuHCl when crush-and soaking acrylamide gel slabs can denature DNA. I normally use water or the Maniatis buffer, but having 5M GuHCl around would help...

02 March 2018 7,428 3 View

Does anyone Happen to know what BioRad's iProof PCR enzyme is (beyond what's in the catalog description)?

09 October 2016 1,608 2 View

How to control pH and composition of PEG solutions?

Hi all, I am trying to concentrate a DNA sample using dialysis against 50% PEG, through a 10K cut-off membrane. I made a 50% solution of PEG20K supplemented with 10mM Tris (target pH=7.8), and...

08 September 2016 3,873 10 View

An affordable and reliable thermal cycler?

02 March 2016 3,750 5 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

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 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

Samples to study circular dichroism (chirality)?

I want to build a low-cost polarimeter to study circular dichroism (chirality). What type of samples can I use? I am looking for samples that can be found outside the laboratory.

01 March 2021 9,101 3 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

How to accurately quantify chitin?

Hi all, I have been working on the quantification of chitin in insect samples and feed for a while but I can notice that the quantification methods (e.g. exposure to HCl, NaOH, and weighing sample...

01 March 2021 1,843 2 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 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