7 replies to this topic

[Muhammad Umer]

Can anybody plz come up with his/her experience using any of the following bisulfite conversion kits

Epitect bisulfite kis (Qiagen)
EZ DNA methylation kit (Zymoresearch)

Also please tell me if anybody has ever used "EZ DNA methylation-Startup kit". How are the results of their PCR components included in this kit?

[RKA]

Muhammad Umer, on Dec 18 2009, 10:19 AM, said:

Can anybody plz come up with his/her experience using any of the following bisulfite conversion kits

Epitect bisulfite kis (Qiagen)
EZ DNA methylation kit (Zymoresearch)

Also please tell me if anybody has ever used "EZ DNA methylation-Startup kit". How are the results of their PCR components included in this kit?

I have used the EZ start up kit. I think it works fi9. In my case i worked with ffpe material, the universal hMLH1 primers provided with the kit to check for the conversion didn't work for my test samples, but worked for control sample. But i did a check with my own primers belonging to the known methylated genes in my study. i think many people have gud experience with EZ kit..Good Luck

[MoB]

Muhammad Umer, on Dec 18 2009, 11:19 AM, said:

Can anybody plz come up with his/her experience using any of the following bisulfite conversion kits

Epitect bisulfite kis (Qiagen)
EZ DNA methylation kit (Zymoresearch)

Also please tell me if anybody has ever used "EZ DNA methylation-Startup kit". How are the results of their PCR components included in this kit?

They are both excellent kits regarding the (for me) most important  parameters bisulfite-conversion, DNA-yield and purity (no PCR inhibiting residuals). Definitely the best bisulfite-conversion kits on the market...

[Muhammad Umer]

thank you both for your valubale comments. I wonder if methylnick can come up with some valuable input

[et2b]

Hi,

in our hands Zymo 96-wells and columns give good conversion rates with the overnight protocol, somehow the cycling protocol does not work that well. I have heard that others have the same problems/observation.
Epitect is very nice for low DNA quantity bisulfite conversion, use the FFPE version of the protocol. Also Epitect does not chew up your DNA as bad as Zymo. However, genome wide studies show that to get more than >99% conversion with epitect you need to do the entire protocol twice on the same sample (Meisner et al. various papers), otherwise it is around 95% only.

Best wishes

[Rainbowz]

็็Hi, I'm Beam

I used Epitect bisulfite kit and i got converted bisulfite DNA more than starting DNA (in concentration)
Why ? How could you suggest me for the point of this problem or this is the normal result that can find in this study.
It is the error rate of conversion and purification or not ?

Help me please ^^

Thank you

[Rainbowz]

็็Hi, I'm Beam

I used Epitect bisulfite kit and i got converted bisulfite DNA more than starting DNA (in concentration)
Why ? How could you suggest me for the point of this problem or this is the normal result that can find in this study.
It is the error rate of conversion and purification or not ?

Help me please ^^

Thank you

Beam_za@hotmail.com

[MoB]

Rainbowz, on 03 May 2011 - 11:53 PM, said:

็็Hi, I'm Beam

I used Epitect bisulfite kit and i got converted bisulfite DNA more than starting DNA (in concentration)
Why ? How could you suggest me for the point of this problem or this is the normal result that can find in this study.
It is the error rate of conversion and purification or not ?

Help me please ^^

Thank you

Beam_za@hotmail.com

How do you quantify your bisulfite converted DNA (UV, real-time PCR)? Maybe your observation is simply caused by the fact that you start with an input volume of 100 µl into the bisulfite conversion but end with an elution volume of 50 µl. If you use carrier RNA and quantify by UV you will definitely measure higher concentrations.

Hope that helps...

MoB

Edited by MoB, 06 May 2011 - 12:50 AM.