Author: He, Biao; Li, Zuosheng; Yang, Fanli; Zheng, Junfeng; Feng, Ye; Guo, Huancheng; Li, Yingying; Wang, Yiyin; Su, Nan; Zhang, Fuqiang; Fan, Quanshui; Tu, Changchun
Title: Virome Profiling of Bats from Myanmar by Metagenomic Analysis of Tissue Samples Reveals More Novel Mammalian Viruses Document date: 2013_4_22
ID: 04d0koah_10
Snippet: To synthesize dscDNA, a 39-59exo -Klenow fragment (5 U; New England Biolabs, Beijing, China) was added to the cDNA mixture and barcode primers, then incubated at 37uC for 60 min, after which the enzyme was inactivated at 75uC for 10 min. To remove phosphates and free single-stranded bases in the dscDNA reaction, 2 U shrimp alkaline phosphatase (SAP, TaKaRa) and 2.5 U exonuclease I (TaKaRa) were added to the dscDNA reaction mixture along with 106S.....
Document: To synthesize dscDNA, a 39-59exo -Klenow fragment (5 U; New England Biolabs, Beijing, China) was added to the cDNA mixture and barcode primers, then incubated at 37uC for 60 min, after which the enzyme was inactivated at 75uC for 10 min. To remove phosphates and free single-stranded bases in the dscDNA reaction, 2 U shrimp alkaline phosphatase (SAP, TaKaRa) and 2.5 U exonuclease I (TaKaRa) were added to the dscDNA reaction mixture along with 106SAP buffer and ddH 2 O to a final volume of 50 ml, then incubated at 37uC for 60 min and inactivated at 75uC for 10 min. Sequence-independent Single Primer Amplification (SISPA) and Purification of PCR Products
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