Clontech
Terra PCR Direct Polymerase Mix
639271
We switched our mouse genotyping PCR system from promega Gotaq flexi to Clontech Terra direct PCR. The new clontech kit can use mouse tissue directly for PCR and does not require optimization. It always give specific DNA band and can amplify high GC content fragment easily.
Mouse genotyping
1mm mouse tail
1) incubate mouse tail with 180 ul 50 mM NaOH for 10mins at 95C 2) Directly take 1ul crude tissue extract in 20 ul PCR system (Kit suggests using 20ul of 1M Tris-HCl ph8.0 to neutralize the extraction, we avoid this step). 3) Following instruction, PCR amplify 33-35 cycles and then take 1/4 for gel analysis.
Kit suggests 50 ul PCR system and include 800 reactions. We found 20ul, or even 10 ul PCR works very good in the lab. Generally I use 10 ul for my mouse genotyping, which means one kit can run 4000 PCR reactions.
Simple handing and it always give me specific DNA bands.
None
Quick, robust and reproducible results
It can only be used for genotyping, not for cloning
Save time, avoid optimization and amplify very difficult DNA fragments