2 replies to this topic

[deilepaita]

Hello there, I hoppe that someone could help me.

I´ve a problem with my real time PCR Rotor Gene 6000.

When I use Qiagen SYBR Green Master Mix, the machine works and it read the fluorescense; but when I use Fermentas SYBR Green Master Mix, this machine can not read the fluorescense. It seems to be lower than Qiagen Master Mix...

I´ve tried to change the gain (manually and automatically) for these master mix too, but still doesn´t work with Fermentas SYBR Green.

I´ve changed the master mix three times, but still doesn´t work.

I don´t know what it is... Why did Qiagen SYBR Green work and Fermentas didn´t?  

Thank you for your help.


Dessy

Edited by deilepaita, 17 November 2009 - 06:25 AM.

[BioMiha]

deilepaita, on Nov 17 2009, 03:22 PM, said:

Hello there, I hoppe that someone could help me.

I´ve a problem with my real time PCR Rotor Gene 6000.

When I use Qiagen SYBR Green Master Mix, the machine works and it read the fluorescense; but when I use Fermentas SYBR Green Master Mix, this machine can not read the fluorescense. It seems to be lower than Qiagen Master Mix...

I´ve tried to change the gain (manually and automatically) for these master mix too, but still doesn´t work with Fermentas SYBR Green.

I´ve changed the master mix three times, but still doesn´t work.

I don´t know what it is... Why did Qiagen SYBR Green work and Fermentas didn´t?  

Thank you for your help.


Dessy

I am attaching a link to a website from our supplier of Fermentas products.
http://www.majbert.c...licnih-napravah
Disregard the fact that it is in Slovene (perhaps you can find this table in English on the Fermentas website). There is a table showing the relative usefulness of different Fermentas qPCR kits with different qPCR mashines. * means good (in reality poor) and *** is very good (in reality doable). I don't know what specific master mix you are using, but you can check how it holds up for your mashine and figure out why other kits are better suitable for that mashine. That should give you a clue, what to look out for.
Hope it helps,
Cheers, Miha

[deilepaita]

BioMiha, on Nov 18 2009, 05:41 AM, said:

deilepaita, on Nov 17 2009, 03:22 PM, said:

Hello there, I hoppe that someone could help me.

I´ve a problem with my real time PCR Rotor Gene 6000.

When I use Qiagen SYBR Green Master Mix, the machine works and it read the fluorescense; but when I use Fermentas SYBR Green Master Mix, this machine can not read the fluorescense. It seems to be lower than Qiagen Master Mix...

I´ve tried to change the gain (manually and automatically) for these master mix too, but still doesn´t work with Fermentas SYBR Green.

I´ve changed the master mix three times, but still doesn´t work.

I don´t know what it is... Why did Qiagen SYBR Green work and Fermentas didn´t?  

Thank you for your help.


Dessy

I am attaching a link to a website from our supplier of Fermentas products.
http://www.majbert.c...licnih-napravah
Disregard the fact that it is in Slovene (perhaps you can find this table in English on the Fermentas website). There is a table showing the relative usefulness of different Fermentas qPCR kits with different qPCR mashines. * means good (in reality poor) and *** is very good (in reality doable). I don't know what specific master mix you are using, but you can check how it holds up for your mashine and figure out why other kits are better suitable for that mashine. That should give you a clue, what to look out for.
Hope it helps,
Cheers, Miha

Thank you very much!!!

I think you´re right Miha.

I was using the Maxima™ SYBR Green/ROX qPCR Master Mix (2X) K0221 in the Rotor Gene 6000, and the Qiagen Master Mix doesn´t have ROX. Maybe I should use the Maxima™ SYBR Green qPCR Master Mix (2X), ROX Solution provided in another tube K0251 OR K0252.



Dessy