GAPDH rodent kit (Applied biosystems)

[Patrick]

Hi,

Is there somebody who has experience with using the GAPDH rodent kit (Aplied Biosystems Cat No. 4308313) and the One step RT-PCR mastermix (Applied Biosystems Cat No. 4309169). Up till know we have never observed any amplicon formed.

At first we always check the reaction on a 2.5% agarose gel and we have never seen a product. However when this combination of kits is used in qPCR a Ct value can be found. However I assume that GAPDH as a household gene should also give a fragment on a gel.

The product specialist of applied biosystems confirmed my finding but they indicated that it should be possible to use this kit wiithout confirmation on a agarose gel.

My question is does someone also works with these kits and have they also have problems and am I a litte bit sceptical if I want to confirm my CT value with an product of the expected size on a 2.5 agarose gel