Sigma-Aldrich
Extract-N-Amp Tissue PCR Kit
XNAT2
Standard genotyping protocols are laborious and time consuming. It involves digestion of minced tail overnight followed by DNA precipitation using isopropanol or phenol-chloroform. If you want an easy and quick method, Sigma's Extract-N-Amp TIssue PCR kit can become your choice if you are ready to spend a few trial for optimization. Then genotyping is a few hours work on the side.
Mouse tail DNA extraction and genotyping
Mouse tail biopsy, Extract-N-Amp Tissue PCR Kit and primers
Collect 4-5 mm tail biopsy following all IACUC guidelines, cut into small pieces using scalpel and forceps, digest it in 100 uL extraction solution and 25 uL preparation solution and incubate at 55C for 20-30 min in a shaking block. Heat inactivate digestion mix at 95C for 3 min. Use 5-10 uL inactivated digest for PCR. Optimize for digestion time and temperature before taking up large batch.
Tail tips from young mice are easy to digest and fresh tissue digests faster than frozen specimens.
We never had failure with samples. PCR resulted in sufficient amplification in 30 cycles. All steps could be completed in 3 hours.
N/A
Easy and quick.
Initial optimization necessary for digestion. Some times PCR trouble shooting was needed.
Saves time if you are ready to spend some for optimization.