We selected 5 kinds of genes randomly and synthesized each Forward or Reverse Primer with Trityl-ON.
Thereafter, we underwent cartridge purification and obtained solution of each primer, then we prepared samples of primer solution containing Primer Stabilizer(+) and Primer Stabilizer(-).
We performed preservative test for those samples under condition of 4 ℃, 25 ℃ and 37 ℃ and compared time course of preservation stability by molecular weight analysis of MALDI TOF/MS.
Followings are data of sample primers.
< Result >
Following are representative examples of MALDI TOF/MF molecular weight analysis.
Primer Stabilizer(-) 25 ℃ Gene A /Forward Primer 1 month(Left), 7 month(Right)
Primer Stabilizer(-) 25 ℃ Gene A /Reverse Primer 1 month(Left), 7 month(Right)
Preservative Test at 37 ℃ Gene E、Forward Primer
Primer Stabilizer(-) 1 week(Left), 8 week(Right)
Primer Stabilizer(+) 1 week(Left), 8 week(Right)
Basic line will start fluctuating irregularly a little from half a year(data not shown)
Apurinic Acid increased 2-fold in one month and increased manyfold after 4 months, also irregular fluctuation of basic line has occurred significantly.
Apurinic Acid increased 2 - 3 fold in one week and increased manyfold in one month, also irregular fluctuation of basic line has occurred significantly.
In general, it is recommended to store primers for in vitro diagnostics in freezer at -20℃ after use, however it was revealed that addition of Primer Stabilizer to the Primer Solution would suppress decomposition of primers even under stress preservative test at 37℃.
< Discussion >
In usual primer solutions, substance to promote cleavage of primers will be generated in the course of time. One of them is oxygen radical. This Primer Stabilizer is considered to function as a scavenger for oxygen radical and as a result, to suppress cleavage of primers.