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Hi there,
Does anyone know what are the enzymes of the Adaptase in Swift (now IDT) ssDNA Library Prep Kit?
The manufacturer claimed that it used Adaptase technology which adds a low complexity polynucleotides tail with a median length of 8 bases to the 3' end of each single-strand DNA fragment(see attachment picture 1). And in a Chinese manufacturer's manual with similar technology, they claimed that it adds poly-dCTP to the 3' end(see attachment picture 2). During the Adaptase step, a ligation reaction is performed at 37℃ for 5 min then 95℃ for 2 min. Obviously, the ligation reaction happens at 37℃, and enzymes are inactivated at 95℃.
I think the poly-C sequence provides higher Tm for splint ligation so the ligation actually is a double-strand ligation. However, what are the enzymes in the ligation? T4 DNA ligase usually loses activity at 37℃.
Am I right? Does anyone know the ligation mechanism of the Adaptase?
Best
Yang
Does anyone know what are the enzymes of the Adaptase in Swift (now IDT) ssDNA Library Prep Kit?
The manufacturer claimed that it used Adaptase technology which adds a low complexity polynucleotides tail with a median length of 8 bases to the 3' end of each single-strand DNA fragment(see attachment picture 1). And in a Chinese manufacturer's manual with similar technology, they claimed that it adds poly-dCTP to the 3' end(see attachment picture 2). During the Adaptase step, a ligation reaction is performed at 37℃ for 5 min then 95℃ for 2 min. Obviously, the ligation reaction happens at 37℃, and enzymes are inactivated at 95℃.
I think the poly-C sequence provides higher Tm for splint ligation so the ligation actually is a double-strand ligation. However, what are the enzymes in the ligation? T4 DNA ligase usually loses activity at 37℃.
Am I right? Does anyone know the ligation mechanism of the Adaptase?
Best
Yang
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