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primer design@ buy?


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#1 hidayahcellculture

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Posted 16 January 2014 - 07:54 AM

CAN WE JUST BUY THE PRIMER INSTEAD OF DESIGN IT?
for example, in this case:

RNA extraction and Reverse transcription-polymerase chain reaction (RT-PCR)

Total mRNA will be isolated from culture supernatants with Rneasy Plus Mini Kit (Qiagen, UK) according to the manufacturer’s instructions prior to conversion to cDNA and will be stored at -20ºC. 

The amount of mRNA will be determined using Quanti-Fast SYBR Green RT-PCR kit (Qiagen, GmbH) with a validated primer set from Qiagen specific for osteogenic markers (runx2, osterix, osteopontin, bone sialoprotein, BMP-2 and IGF-1). 
The threshold cycle (Ct), the cycle number at which the amount of amplified gene of interest reached a fixed threshold, will be determined to the manufacturer’s instructions. 
Relative expression of the RT-PCR product will be calculated using the comparative 2_ΔΔCt method. The endogenous control glyceraldehyde-3-phosphate dehydrogenase mRNA will be used for normalization. 

The fold difference will be then determined by normalizing all values to the mean of the relative expression for the control group without treatment. 

Finally, the PCR product will be electrophoresed on ethidium bromide-stained agarose gels and visualized under UV illumination. 

primer set from Qiagen specific for osteogenic markers (runx2, osterix, osteopontin, bone sialoprotein, BMP-2 and IGF-1).



#2 bob1

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Posted 16 January 2014 - 11:49 AM

Yes, if they have been published and validated before, it makes your life a bit easier.  You don't need to re-invent the wheel for your research.



#3 hidayahcellculture

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Posted 16 January 2014 - 02:12 PM

if that case, thank you MR moderator..






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