Protein Pellet Resuspension with how much DTT?

I have extracted my protein using isopropanol from Tri-reagent LS extracted cell culture. I found the pellet is difficult to dissolve in 1% SDS. I have read through some articles saying that...

16 January 2017 8,549 3 View

Do I need to standardise the exposure time of the target protein & housekeeping protein with ECL?

I can see my target protein band clearly around 333 seconds but for my housekeeping protein, I can see it clearly around 10 seconds with total protein loading is 40ug. May I know do I need...

31 October 2016 5,647 4 View

Why human serum used as 1st antibody always cause high background in western blot?

My western blot protocol was : Sample:normal human epidermal extract, 25 ug/lane 7.5%PAGE, semi-dry transfer to nitrocellulose membranes. Blocking buffer:5% skim milk in TBST(0.5% TWEEN 20) 1st...

18 January 2016 8,577 7 View

How can I analyse gene expression time course study by qPCR?

I am doing time course study for gene expression in cell culture. I am doing 0, 1, 2, 4, 8, 16, 24 hours. Each time point I have treated and untreated (control). I should normalised my individual...

14 December 2015 5,249 6 View

Do gene expression real time RT-PCR study have a big biological replicate differences?

I am doing real time RT-PCR of human cell line. May I know if the gene expression in different biological replicates will be different or almost the same?

26 November 2015 8,282 3 View

Can I optimize dose and time in different cell lines?

I am doing dose and time response in animal cell culture. Do I have to optimize the dose and time in one particular cell line first, then use the optimized dose for another 2 cell lines? Or, do I...

15 November 2014 6,564 2 View

Will the secondary structure of PCR products affect the mobility in agarose gel electrophoresis?

12 September 2014 2,943 6 View

Abrnomal Melt Peak qRT-PCR

There is a peak next to the 80C melt peak. May I know what is the problem? Primer dimers?

14 May 2014 9,871 2 View

How do I reduce smear on a qRT-PCR melt curve?

I obtained this melt curve from qRT-PCR (one Step RT-PCR). The peak is so broad and when I run the gel, there was primer dimer smear below the desired product. How do I reduce this primer dimer...

05 May 2014 5,213 42 View

Can anyone help with transforming Qiagen SYBR Green PCR (2 steps real time PCR) to Qiagen SYBR Green RT-PCR (qRT-PCR)?

I have optimized the cycling condition using 2 steps Quantifast SYBR Green from Qiagen but when I try out one step Quantifast SYBR Green, I could not get the desired PCR product. In 2 steps real...

24 April 2014 1,104 3 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

Resistor definition in Piezoelectric bimorph with ANSYS workbench

Hello, good day! I am trying to simulate similar bimorph piezoelectric harvester with below commands. However, the output power doesn't change with resistor value. Please help to find the reason. ET,10,CIRCU94,0 !SET UP RESISTOR R,1,100000 !RESISTANCE VALUE TYPE,10 !SET ELEMENT TYPE E,1,2 !CREATE RESISTOR BETWEEN NODES 1 AND 2 Thanks in advance Regards,

02 March 2021 0 0 View

No title

02 March 2021 3,060 3 View

Why are the velocities so high in my Boussinesq approximation Fluent simulation?

Hello, I have a mechanism where air rises due to convective flows. In this simulation, nothing is driving the air except this. I understand that the Boussinesq approximation only works for small...

02 March 2021 4,910 3 View

What is the characteristic length to determine the Reynolds number for the scattered turbulent fluid flow?

Dear all, I just saw this example in FB on the difference between laminar flow and turbulent flow. The flow from a pipe hits the sink and scattered. The flow from the pipe is characterized as...

02 March 2021 1,999 5 View