Reconditioning process validation - 1.5 mL tubes used for PCR
New and reconditioned 1.5 mL tubes were used to ensure they maintain their integrity through the reconditioning process.
PCR reaction mix for pGFP-actine was made in 10 different 1.5 mL tubes. The content of 5 of them is then transferred in 5 PCR tubes. After the PCR, loading buffer is added to the PCR tubes and samples are placed in the first wells n°2-6 of an agarose gel.
The ten 1.5mL tubes are cleaned and dried through our reconditioning process. Five of them are then used again to prepare a PCR reaction mix for pGFP-actine and the other five are used as negative controls for the same reaction (containing all reagents except the pGFP-actine). Their content is then transferred in 10 PCR tubes. After the PCR, loading buffer is added to the PCR tubes and samples are placed in wells n°7-11 of an agarose gel (process with pGFP-actine) and wells n°13-17 (negative controls with reconditioned tubes). Well n°12 contains a negative control prepared in a new 1.5 mL tube and acts as a true negative control. Well n°1 contains the molecular weight marker.
This experiment allowed us to test for residual carryover of DNA.
Results
All reconditioned tubes led to no signal in their respective wells, indicating that they contained no DNA carryover.
Conclusion
Our reconditioning process operated in a way that will allow the use of washed and dried consumables to be used multiple times in the Molecular Biology Department.
