Trehalose as a good candidate for enriching full-length cDNAs in cDNA library construction.
It has been reported that the disaccharide trehalose is capable of increasing the thermostability and thermoactivity of reverse transcriptase, and therefore improving the length of cDNA synthesis. However, no test has been done on how the disaccharide trehalose performs in the context of the entire cDNA synthesis processes, or whether it can seamlessly integrate into the commercially available cDNA synthesis kit. In this report, we optimized a protocol to incorporate trehalose in the Stratagene's cDNA library construction kit in order to demonstrate great improvement in cDNA's length (average length of 1.8 kb in the trehalose group versus 1.0 kb in the control). Sequence analysis of the cDNA clones showed that the addition of trehalose did not increase the error rate of the RT products but greatly increase the quantity of full-length in cDNA library.
Chen
Lei
L
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China.
Cao
Lixue
L
Zhou
Longhai
L
Jing
Yudong
Y
Chen
Zuozhou
Z
Deng
Cheng
C
Shen
Yu
Y
Chen
Liangbiao
L
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D005399:Fishes / Q000235:genetics*; D015723:Gene Library; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014199:Trehalose / Q000737:chemistry*
D018076:DNA, Complementary; D014199:Trehalose