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Illumina cDNA library preparation and sequencing: Approximately 30 g of total RNA with a concentration of [greater than or equal to] 250 ng/ L was used for cDNA synthesis and Solexa sequencing.
Polymerase chain reactions for familial analysis and cDNA library generation were carried out in a final volume of 20 [micro]l using the high-fidelity cloned Phusion polymerase (New England Biolabs), using appropriate buffers as the instructions indicated.
We sequenced unamplified poly(A)-tailed viral RNA using rapid cDNA library preparation coupled with real-time data analysis to determine its potential application for pathogen genomic characterization.
Among these immunoreactive autoantigens, there are 2 proteins; in particular, Zuotin-related factor 1 (ZRF1) and R motif-containing protein 1 (KRR1) (clones KYMBC 29-88-1 and KY-MBC 18-53-1) were identified during the screening of cDNA library from medullary breast carcinoma.
cDNA library construction and high-throughput sequencing
cDNA coding for human CD19 was first cloned and described by Tedder et al from human tonsil cDNA library (3).
For their experiments young developing ovaries were the organ of choice for total RNA isolation for cDNA library generation.
Novagen's T7Select Phage Display vector for construction of cDNA library.
6] sequence reads 50 bp in length (Table 1) (see cDNA Library Preparation in the online Data Supplement for a description of salivary cDNA library preparation).
arrayed Strorgvlocetrotus purpuratus cDNA clone Spl170 5', similar to Actin, mKNA sequence gi|139309108 AM569285 RZPD 2716 32092 MPMGp1173 Paracentrotus lividus CASPLU Paracentrotus lividus cDNA clone MPMGp1173C12115Q 5', mRNA sequence gi|109400265 132678021 Sea Urchin 1793 38193 Mesentary cDNA library Strongylocentrouce purpuratus cDNA done MSP-2L19 5', mRNA sequence gi|24901528 RN.
Third, a relatively large quantity of mRNA is required to obtain a cDNA library of even moderate size because of overall low cloning efficiencies.
In the present study, we constructed subtractive cDNA library with urine exfoliated cells to screen differentially expressed genes in patients with BTCC and to identify some urine markers for BTCC.