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

Can I use breaking series as a regressor in a VAR model?

Context: I have two series, price and sales. Sales is mean-reverting stationery but price is stationary only after controlling for an intercept break. I want to set up a 2-equation VAR model...

14 January 2021 6,824 12 View

Does the use of Tissue Level implants reduce the risk of Peri-implantitis, as opposed to using bone level?

12 December 2020 6,007 6 View

Will the plasmid without SV40 Ori have a quite low expression level in 293T?

SV40 Ori helps plasmid to replicate in 293T cells. If I use a plasmid which doesn't have a SV40 Ori, will the expression level be very low?

10 December 2020 1,751 2 View

How to calculate the normalized frequency V of photonic crystal fiber using COMSOL?In particular, how to calculate NEFF and nFSM using COMSOL?

24 November 2020 967 2 View

HOW to do an intention-to-treat analysis(ITTA)?

I mean, how do I put the dropped out participants' outcome (Missing value) in my analysis? Although I have already known what's ITTA meaning, I still don't know how to do it. Which methods should...

23 November 2020 9,982 5 View

Cloudy brain samples in sucrose solution?

I have some samples I left in sucrose for a couple of weeks after perfusion and was wondering if these samples are still salvageable? I perfused the brain, then they were fixed in 4% pfa for 24...

15 November 2020 1,728 2 View

Does pEGFP plasmid have G418 resistant?

Instruction from pEGFP: The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected usingG418 ....

20 October 2020 4,662 2 View

The solubility of ethinyl estradiol (EE2) in methanol?

What is the solubility of ethinyl estradiol (EE2) in methanol? In the experiment, methanol was used to help dissolve, what is the reason for the flocs appearing after the storage solution is added...

20 September 2020 4,055 1 View

How to compare methylation status between different cell lines?

As in the title. I got the methylation status of several genes in different cell lines by bisulfite sequencing. More specifically, I have the sequencing data from the amplicons of these genes...

30 August 2020 6,020 2 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

What analysis to choose with a large neste dataset and clearly skewed or kurtotic distributions?

Hello! I have a dataset of n=3000 nested within 8 countries with approximately 200 or 400 responses in each country. I originally planned to perform multilevel modelling with 4 dependent variables...

02 March 2021 6,865 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

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