1, 2 Ligation of adaptors is not only time consuming but also a low efficiency process that requires micrograms of nucleic acid inputs. Current methods for MPS libraries construction utilize either RNA or DNA adaptors ligation to the 5′- and 3′- ends of the target RNA or DNA molecules.
Massive parallel sequencing (MPS) of nucleic acids requires the preparation of amplified libraries where the DNA region of interest is located between known 5′- and 3′- terminal sequences. The approach for DNA library preparation from minimal and fragmented input described here will find broad application in diverse research areas such as translational medicine including therapy monitoring, prediction, prognosis and early detection of various human disorders and will permit high-throughput DNA sequencing from previously inaccessible material such as minute forensic and archeological samples. To exemplify the capacity of the technique, we have generated and sequenced DNA libraries from hundred-picogram amounts of circulating nucleic acids isolated from human blood plasma, one nanogram of mRNA-enriched total RNA from cultured cells and few nanograms of bisulfite-converted DNA. Furthermore, those DNA libraries are strand-specific, can be prepared within 2–3 h and do not require preliminary sample amplification steps. We have established a fast, cheap and highly efficient protocol called ‘Capture and Amplification by Tailing and Switching’ (CATS) to directly generate ready-to-sequence libraries for MPS from nanogram and picogram quantities of both DNA and RNA. This together with high cost and time requirements impedes many important applications of MPS in medicine and fundamental science. However, sequencing of trace amounts of DNA or RNA still remains a major challenge, especially for degraded nucleic acids like circulating DNA. The app uses one or more features on the device for determining location, such as GPS location, network location, or cell location.#The app uses precise location coordinates obtained from a Global Positioning System (GPS) receiver on the device.#The app uses coarse location coordinates obtained from a network-based geolocation system supported on the device.#other.Massive parallel sequencing (MPS) technologies have paved the way into new areas of research including individualized medicine.
The app uses the Global System for Mobile Communications (GSM) telephony radio system. Allows using PowerManager WakeLocks to keep processor from sleeping or screen from dimming.Īllows an application to request installing packages.Īllows applications to open network sockets.Īllows applications to access information about networks.Īllows an application to write to external storage.Īllows an application to read from external storage.Īllows read only access to phone state, including the phone number of the device, current cellular network information, the status of any ongoing calls, and a list of any PhoneAccounts registered on the device.Īllows an app to access precise location.Īllows an app to access approximate location.ġ20, 160, 213, 240, 320, 480, 640, 65534, 65535
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