Differences between MSP and BSP? - (Feb/28/2005 )
running at high voltage and getting that result doesn't really matter your size range is fine for IP
N
Greetings fellow MSP and BSP'ers!
All of your postings on this board have been most helpful! And I thank you all for that.
I understand the concept between the differences in the primers for MSP vs BSP. However, if you're gonna do PCR and get a band regardless, then can't you sequence the band in MSP too? Or has the DNA been treated differently in MSP than BSP so that sequencing cannot be performed in MSP? 
Thanks
Do you have any recommendations as to what company has the easiest to use kit for methylation studies?
Thanks
The modification step is the same for MSP and BSP. If you want to do sequencing, only sequence BSP products. Sequencing MSP products doesn't make sense.
From its protocol, it seems to me EZ DNA Methylation Kit is very simple but I have not used it myself.
The modification step is the same for MSP and BSP. If you want to do sequencing, only sequence BSP products. Sequencing MSP products doesn't make sense.
But aren't MSP studies not as credible as BSP studies? Sequencing your bands will give THE most conclusive evidence? no? That's why I was wondering about sequencing MSP products.
MSP tests only CpGs within the primer set, BSP tests all CpGs within the amplicon (a whole lot more potentially) so I wouldn't say BSP is more crdeible, but that BSP assays more sites and MSP, MSP is a quick way of determining methylation status without the need to goto sequencing, and you shouldn't be sequencing your MSP products as these results, as you said, are meaningless.
Nick
Hi everybody, MSP sucks hard. 
It overestimates effects 
, is not really quantitative and measures only 1 CpG and then people say that a entire promoter is "hypermethylated" or "hypomethylated"....and than that is used for medical research..... 
to convince yourselves:
read: J Nutr. 2005 Jun;135(6):1382-6 from Lillycrop et al
and be "convinced" 
than read: and start thinking about the validity of MSP in every case...knowing that bisulfite treatment if done properly still gives a 2% batch effect easily when using pyrosequencing or Epityper.
Br J Nutr. 2008 Jan 11 : lillycrop et al. Please not that the samples they use are the same as the 2005 study.....and sample size is small....
Happy thinking people.
I assume by BSP you mean bisulfite PCR and sequencing. If so, the advantages of this over MSP are:
A greater number of CpG residues analysed for methylation when compared with MSP (MSP is only one and that is identified with a methylation specific primer). Bisulfite sequencing looks at every CpG residue across the whole amplicon.
I would also say that primer optimisation is crucial for an MSP assay to work, it is not so crucial for BSP because you will be sequencing through the amplicon anyway.
MSP has it advantages in that it is far more quicker than BSP in terms of hands-on lab time.
hope this helps. I would be interested to see what other people think about this also.
Cheers
nick
Hi, nick
If I use the MSP to compare the methylation intensity induced by drug treatment, then further use the BSP to show the sequencing result
Do this BSP product need cover the MSP product sequence
Thanks
Anne