In PCR, why do the salts differ for particular polymerase enzymes like Mgcl2 for Taq and MgSo4 for Kod polymerase?

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What if top fiber concrete strain exceeds the ultimate strain when fixing the yield strain in steel i.e to calculate yield moment in circular column?

I have prepared an excel sheet to calculate yield moment and curvature for a hollow circular section. In which Depth of Neutral Axis is fixed using "GOAK SEEK" function. Axial Force is input of...

22 November 2020 4,632 3 View

Should I perform transcardial perfusion by PBS before lung isolation for RNA expression analysis?

Hey, I am planning to dissect rats and isolate Lung tissue samples to be stored in RNA stabilizer for later analysis. - Should I perform trandcardial perfusion by PBS ? - Is it necessary to...

21 September 2020 1,822 5 View

ELISA of isomer of glycolytic intermediate 2,3 DPG ?

Hello, I recently performed Kit based ELISA for 2,3 DPG on plasma samples of control and hypoxia treated rats. [CAT. NO. E1562Ra, Bioassay technology laboratory] However, the results I obtained...

07 September 2020 3,270 2 View

Can I use one facet of a multidimensional latent variable questinaairre/scale?

I wish to use self-monitoring scale developed by Richard D. Lennox and Raymond N. Wolfe ("Revision of self-monitoring"). It is a 2 factor structure scale with total 13 items- (1) ability to modify...

17 May 2020 8,027 3 View

On column TEV cleavage?

Hi, I have cloned my protein with TEV protease site (His tag-TEV site-linker-protein). I purify my protein with Ni-NTA and then do on column TEV cleavage (overnight). I hardly see any TEV...

10 March 2020 1,205 3 View

How to get a good SDS page gel for 16KDa protein?

I want to see a 16kDa band of my protein in SDS PAGE. Whenever i run the gel, the band in the upper part of resolving gel were very sharp and clear however the lower part where i supposed to get...

11 February 2020 7,749 2 View

What is current state of machine learning in the field of condition monitoring?

I am looking for the possible implementation of ML tools for electrical machines. If any of the approach has even been used in the industry?

28 January 2020 8,165 3 View

Any Suggestions to rectify negative intercept in HPLC?

Drug is Sodium citrate, diluting drug in de-ionized water. Using a C18 column. the concentration range of 1000-3000microgram to lower range of 25-300microgram. Thanks!

19 September 2019 7,315 3 View

What length of long non coding RNA (lncRNA) should be used for its overexpression in plants?

Hi all, I am planning to over-express a lncRNA (under 35S) in Arabidopsis. Will the presence of 20-30 bp sequences upstream and downstream of lncRNA sequence affect its over-expression? I...

15 August 2019 6,720 1 View

What would be the mobile phase for quantifying sodium citrate using HPLC method?

I have to determine concentration of sodium citrate using HPLC method. Previous literature's used potassium phosphate buffer of pH 2.5 as mobile phase (isocratic). As there is risk of clogging the...

31 July 2019 1,101 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

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

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

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View