Shin Wei Tie

11 Questions 29 Answers 0 Followers

Questions related from Shin Wei Tie

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...

17 January 2017 8,566 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...

01 November 2016 5,662 4 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...

15 December 2015 5,264 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?

27 November 2015 8,296 3 View

Can I use different final primer concentrations for target and reference genes in qRT-PCR?

I used 100nM and 500nM final concentration for my target gene & reference gene respestively in real time RT-PCR. May I know if this is acceptable?

18 April 2015 220 5 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...

16 November 2014 6,581 2 View

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

If I check the amplicon through MFold from IDT website and find that the Tm of the amplicons secondary structure is around 40C, will it affect the mobility of the products in agarose gel...

13 September 2014 2,963 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?

15 May 2014 9,886 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...

06 May 2014 5,231 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...

25 April 2014 1,118 3 View

How to prepare a complete F12K medium for HUVEC?

I have one bottle containing 500ml F12K. In order to make a complete medium, I have to add in 0.1mg/ml heparin and 0.05mg/ml ECGS. How do I prepare heparin and ECGS?

06 May 2013 3,437 3 View