BioMicroCenter:DNA Sample Preparation

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(New page: --Under construction-- DNA sample preparation is performed on the SPRI-te Nucleic Acid Extractor from Beckman Coulter Genomics, followed by PCR enrichment. The SPRI-te automates end repa...)
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DNA sample preparation is performed on the SPRI-te Nucleic Acid Extractor from Beckman Coulter Genomics, followed by PCR enrichment.  The SPRI-te automates end repair, A addition, illumina adapter ligation, and size selection saving multiple hours of hands on time.  Samples are then completed by PCR, with barcoding available during this step if requested.   
DNA sample preparation is performed on the SPRI-te Nucleic Acid Extractor from Beckman Coulter Genomics, followed by PCR enrichment.  The SPRI-te automates end repair, A addition, illumina adapter ligation, and size selection saving multiple hours of hands on time.  Samples are then completed by PCR, with barcoding available during this step if requested.   
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[[Image:DNA_prep_1.png|center]]
[[Image:DNA_prep_1.png|center]]
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In order to test performance of the SPRI-te the same ChIP-seq sample was prepared for illumina sequencing by an experienced technician and on the SPRI-te.  The automated SPRI-te sample preparation outperformed the sample prepared by hand; number of unique reads and significant peaks was doubled in the SPRI-te samples.  
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[[Image:DNA_prep_2.png|right]]
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[[Image:DNA_prep_2.png|right]]In order to test performance of the SPRI-te the same ChIP-seq sample was prepared for illumina sequencing by an experienced technician and on the SPRI-te.  The automated SPRI-te sample preparation outperformed the sample prepared by hand; number of unique reads and significant peaks was doubled in the SPRI-te samples.  
SPRI-te prepared samples have produced good quality data even below manufactures recommend input levels.  Varying amounts (200ng, 50, 12.5, 3.1, 0.8 and 0.2) of 50bp ladder from New England Biolabs was prepared on the SPRI-te resulting in similar yields and nearly identical traces when analyzed on the bioanalyzer.  
SPRI-te prepared samples have produced good quality data even below manufactures recommend input levels.  Varying amounts (200ng, 50, 12.5, 3.1, 0.8 and 0.2) of 50bp ladder from New England Biolabs was prepared on the SPRI-te resulting in similar yields and nearly identical traces when analyzed on the bioanalyzer.  
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[[Image:DNA_prep_3.png]]  
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[[Image:DNA_prep_3.png]] [[Image:DNA_prep_4.png]]
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[[Image:DNA_prep_4.png]]
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Revision as of 15:01, 18 July 2011

--Under construction--

DNA sample preparation is performed on the SPRI-te Nucleic Acid Extractor from Beckman Coulter Genomics, followed by PCR enrichment. The SPRI-te automates end repair, A addition, illumina adapter ligation, and size selection saving multiple hours of hands on time. Samples are then completed by PCR, with barcoding available during this step if requested.


In order to test performance of the SPRI-te the same ChIP-seq sample was prepared for illumina sequencing by an experienced technician and on the SPRI-te. The automated SPRI-te sample preparation outperformed the sample prepared by hand; number of unique reads and significant peaks was doubled in the SPRI-te samples.


SPRI-te prepared samples have produced good quality data even below manufactures recommend input levels. Varying amounts (200ng, 50, 12.5, 3.1, 0.8 and 0.2) of 50bp ladder from New England Biolabs was prepared on the SPRI-te resulting in similar yields and nearly identical traces when analyzed on the bioanalyzer.

Image:DNA_prep_3.png Image:DNA_prep_4.png

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