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- Picelli, S, et al.
(författare)
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Tn5 transposase and tagmentation procedures for massively scaled sequencing projects
- 2014
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Ingår i: Genome research. - : Cold Spring Harbor Laboratory. - 1549-5469 .- 1088-9051. ; 24:12, s. 2033-2040
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Tidskriftsartikel (refereegranskat)abstract
- Massively parallel DNA sequencing of thousands of samples in a single machine-run is now possible, but the preparation of the individual sequencing libraries is expensive and time-consuming. Tagmentation-based library construction, using the Tn5 transposase, is efficient for generating sequencing libraries but currently relies on undisclosed reagents, which severely limits development of novel applications and the execution of large-scale projects. Here, we present simple and robust procedures for Tn5 transposase production and optimized reaction conditions for tagmentation-based sequencing library construction. We further show how molecular crowding agents both modulate library lengths and enable efficient tagmentation from subpicogram amounts of cDNA. The comparison of single-cell RNA-sequencing libraries generated using produced and commercial Tn5 demonstrated equal performances in terms of gene detection and library characteristics. Finally, because naked Tn5 can be annealed to any oligonucleotide of choice, for example, molecular barcodes in single-cell assays or methylated oligonucleotides for bisulfite sequencing, custom Tn5 production and tagmentation enable innovation in sequencing-based applications.
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- Winberg, Hans, et al.
(författare)
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The complication rate after hypospadias repair correlated to preoperative symptoms
- 2014
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Ingår i: Open Journal of Urology. - : Scientific Research Publishing, Inc.. - 2160-5440 .- 2160-5629. ; 4, s. 155-162
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Tidskriftsartikel (refereegranskat)abstract
- Aim: To assess the rate of complications following hypospadias repair in a consecutive series of boys and the correlations of those complications with their preoperative symptoms, degree of hy- pospadias and method of operation. This study was conducted to address the question of whether all boys with all degrees of hypospadias should undergo reconstruction. Methods: This was a prospective cohort study. We included every boy who underwent an operation for the primary repair of hypospadias between January 2011 and April 2014. The median follow-up time was 24 months. The study ended in October 2014. The main outcome measurements were the frequency of postoperative complications and their correlations with the degree of hypospadias, the preo- perative symptoms and the operative intervention performed. Results: Among the 76 boys who underwent operations, 23 had degree 1, 47 had degree 2, and 6 had degree 3 hypospadias. Preo- peratively, 43 of the boys had symptoms that motivated the operation, including stenosis (38), a curvature (10) or both (5). Forty-three boys underwent operations with the MAVIS technique, 28 underwent TIP repair, 1 underwent a Duckett procedure, and 4 underwent Byar two-stage proce- dures. There were complications requiring reoperations including fistulas or ruptures in 26 (34%) boys. There were no significant differences in the rates of complications with surgery, fistulas (P = 0.4775), ruptures (P = 0.2417) or other complications (P = 0.5165) between the groups with or without preoperative symptoms, those with different degrees of hypospadias or those who underwent different operative methods for repair. Conclusions: The complication rate in this series was high. The study was prospective, and no boy was lost during follow-up. Because the complication rate did not correlate with the degree of hypospadias nor the preoperative symptoms, there may be a group of boys with hypospadias without symptoms for whose operations are questionable. The preoperative symptoms should be reported in future reports of the results of hypospadias surgery.
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- Zhang, XN, et al.
(författare)
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An update on viral association of human cancers
- 2013
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Ingår i: Archives of virology. - : Springer Science and Business Media LLC. - 1432-8798 .- 0304-8608. ; 158:7, s. 1433-1443
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Tidskriftsartikel (refereegranskat)
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