NGS Service for Genomic DNA from Genetic Screen done with transOMIC Library Add to Cart
|Quantity:||NGS Profiling Data and Bioinformatics Analysis|
transOMIC Pooled shRNA Library NGS and Deconvolution Service
Need help deconvoluting your pooled screen?
Send us your genomic DNA samples for sequencing and analysis.
•Simplify your screen - Sequencing service including library preparation, next generation sequencing and data analysis
•Cost effective - Multiplex sequencing allows analysis of multiple samples in parallel
•Quick turnaround - 4-6 weeks from receipt of samples to delivery of data
•More data reproducibility - Optimized protocols and reagents for shRNA specific sequencing
In addition to providing transOMIC´s highest quality sensor-based shRNA pooled library, we also offer their next generation sequencing analysis and deconvolution services. Provide us with the genomic DNA from the samples following your screen and we will deliver the precise number of sequencing reads of each individual clone within a pooled shRNA sample.
The pooled shRNA screen sequencing and deconvolution service is composed of the following steps:
1. Your genomic DNA samples are evaluated for concentration and quality by Qubit® and agarose gel electrophoresis, respectively.
2. Integrated shRNA are amplified from the genomic DNA using mir30 specific primers while maintaining at least 1000X coverage of each shRNA.
3. Following column clean up and quantitation, low-cycle enrichment is performed using Illumina® adapted and indexed primers. Indexed-amplicon libraries are column purified, quantitated and evaluated by agarose gel electrophoresis.
4. If necessary, magnetic bead purification or gel purification is performed to remove unamplified gDNA or primers.
5. Amplicon libraries are next diluted to a 10nM concentration followed by pooling of the individual samples.
6. The pooled sample is next denatured, diluted and loaded according to HiSeq® protocols and recommendations.
7. FASTQ data files for the 50bp sequencing run are produced.
8. Sequence analysis is performed and counts are provided for each hairpin in each sample.