Validated Protocol for Processing with AmpFlSTR® Identifiler® Direct using Prep-N-Go®
The following protocol has been validated for processing the Bode Buccal DNA Collector with the AmpFlSTR® Identifiler® Direct PCR Amplification Kit.
Validated Protocol for Processing with AmpFlSTR® Identifiler® Direct using Bode PunchPrep
|Remove one 1.2mm punch from the Buccal DNA Collector and place into tube or PCR plate well.|
|Add Bode PunchPrep Solution (Cat No. P09D79) and incubate on a 70oC heat block for 20 minutes.|
|Add 25µl of AmpFlSTR Identifiler reaction mix to the 1.2 mm punch and perform a *26-cycle PCR reaction according to the instructions found in the AmpFlSTR® Identifiler® Direct PCR Amplifcation Kit User Guide. *It is recommended that internal validation and testing regarding the number of PCR cycles is performed by each agency. The number of cycles within this protocol should serve as a recommendation for processing.|
Results from Prep-N-Go™ Buffer/AmpFlSTR® Identifiler® Direct evaluation. 159 samples were processed following the Prep-N-Go™/ Identifiler® Direct procedure.
|Number of Amplification Failures||Number of Homozygote Off-Scale Profiles||1st Pass Success Rate|
|Mixture Profile||Partial Profile||No Profile||Heterozygotes: >75 RFU Homozygotes: > 200 RFU|
AmpFlSTR® and Identifiler® are registered trademarks of Life Technologies Corporation
DNA Profile of a sample processed with Prep-N-Go™ Buffer/AmpFlSTR® Identifiler® Direct