Automated DNA-RNA Capture Protocol

Capture of DNA-RNA complexes was performed using the Bravo configured with the 96LT pipetting head, one low plate pad at position 2, and plate heaters (V&P scientific, San Diego, CA, USA, VP-741BW MICA) at positions 2 and 7. All liquid handling steps used 180-μl disposable tips (Agilent Technologies, catalogue number 08585-002). Reactions were carried out according to manufacturer's specifications in the SureSelect Target Enrichment System Sequencing Platform Library Prep v2.0 (Agilent Technologies, catalogue number G3360-90000). Wash protocols were modified to increase the number of wash iterations while decreasing wash buffer volumes to allow wash steps to take place in microtiter plates. See automated protocol in supplementary material for details (Additional file 14).

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Fisher S., Barry A., Abreu J., Minie B., Nolan J., Delorey T.M., Young G., Fennell T.J., Allen A., Ambrogio L., Berlin A.M., Blumenstiel B., Cibulskis K., Friedrich D., Johnson R., Juhn F., Reilly B., Shammas R., Stalker J., Sykes S.M., Thompson J., Walsh J., Zimmer A., Zwirko Z., Gabriel S., Nicol R, & Nusbaum C. (2011). A scalable, fully automated process for construction of sequence-ready human exome targeted capture libraries. Genome Biology, 12(1), R1.

Publication 2011

Buffer Head Library Mica

Corresponding Organization :

Other organizations : Broad Institute

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Protocol cited in 34 other protocols

Variable analysis

independent variables

Bravo configured with the 96LT pipetting head
Low plate pad at position 2
Plate heaters (V&P scientific, San Diego, CA, USA, VP-741BW MICA) at positions 2 and 7
Wash protocols modified to increase the number of wash iterations while decreasing wash buffer volumes

dependent variables

Capture of DNA-RNA complexes

control variables

All liquid handling steps used 180-μl disposable tips (Agilent Technologies, catalogue number 08585-002)
Reactions were carried out according to manufacturer's specifications in the SureSelect Target Enrichment System Sequencing Platform Library Prep v2.0 (Agilent Technologies, catalogue number G3360-90000)

controls

No positive or negative controls were explicitly mentioned.

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