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Questions related from Shin Wei Tie
I would like to do PCR array on IQ5 instrument. This machine has previously done calibration files. Do I need to do calibration again?
08 August 2017 5,594 1 View
May I know which statistical analysis is suitable in western blot for 3 biological replicates? Using arithmetic means or can I use geometric mean?
07 July 2017 4,022 0 View
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...
01 January 2017 2,734 3 View
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...
12 December 2015 4,669 7 View
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?
11 November 2015 9,507 3 View
I am doing on cell culture and I have done 3 biological replicates (n=3) to get my data. I treated my cells with cytokines with five different doses and is compared to control (untreated). I get...
05 May 2015 6,288 9 View
I have done my 3 plates for 3 biological replicates for my control and the samples. May I know how to combine all these 3 results into one? I know this can be done through CFX manager Gene Study...
05 May 2015 6,662 2 View
I am doing 1 step qRT-PCR using quantifast SYBR RT-PCR and using the recommended protocol by Qiagen. I get pretty narrow peak for positive control (HepG2) but not really for THP-1 induced...
02 February 2015 4,508 2 View
I have designed a set of primers with the amplicon length of 137bp, I have done my qRT-PCR but it is not specific . I have done the sequencing but the result is only showing the 23bp which is the...
12 December 2014 6,152 10 View
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...
11 November 2014 5,547 2 View
Dear All, I used HepG2 as positive control as my GOI is confirmed to be expressed in HepG2. It had a melt peak. But there was one minor peak adjacent to 81C peak. May I know what is the problem?...
09 September 2014 7,930 4 View
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...
09 September 2014 9,540 7 View
I have been trying many times and still failed to give single peak in melt curve. I have designed a new primers to detect all 6 variant of my target gene. Please refer to the blast result as...
08 August 2014 134 11 View
I have a dissociation curve like this as attached. May I know if another peak is primer dimer or another products? I blast my primers, one of the variants (total 6 variants) can only be amplified...
08 August 2014 5,983 18 View
I have a pair of primers which should amplify 427 bps of my gene of interest. But when I did qRT-PCR, there is only around 230-240 bps PCR products being amplified and there was smear when I ran...
07 July 2014 5,851 11 View
There is a peak next to the 80C melt peak. May I know what is the problem? Primer dimers?
05 May 2014 4,839 3 View
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 1,756 43 View
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...
04 April 2014 7,672 4 View
May I know if anyone has done this before and how to setup the protocol in Biorad Icycler real time instrument?
03 March 2014 6,089 5 View
I have run the TD-PCR in Bio-Rad Icycler IQ instrument but I can't get one band while I can get one band in conventional TD-PCR. May I have the advice from the experts here? Thank...
03 March 2014 2,362 6 View
What is the difference if I collect real time data in annealing and extension step? Which steps should I setup the data acquisition for real time touchdown PCR?
03 March 2014 153 7 View
May I know if anyone can provide the hFXR protocol as I can't amplify for Real-Time RT-PCR?
12 December 2013 4,749 0 View
I have done qPCR. There is a minute expression of FXR in THP-1 monocytes (0.001 if compare to HepG2).
10 October 2013 1,140 0 View
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?
05 May 2013 9,419 3 View
