Sample Submission | VCR | University of Nebraska Medical Center

Methylation Analysis
The amount of purified DNA (Proteinase K and RNase A treated) needed for each type of methylation procedure varies according to the type of analysis and the number of genomic regions that need to be analyzed. The following table is a guide you can use to determine the amount of needed DNA, however, we have obtained excellent results from much smaller quantities. If your DNA supply is limited contact us to discuss your possibilities or other feasible approaches.

Procedure	Optimum DNA amount needed
Methylation Specific PCR	200 ng/two primer locations
Bisulfite Sequencing	200 ng/genomic region
Bisulfite Pyrosequencing	200 ng/genomic region
Methyl-sensitive cut counting Genomic Sequencing (MSCC)	2 µg per sample/restriction endonuclease digestion
Methyl CpG Binding Domain - Isolated Genome Sequencing (MiGS)	5 µg per sample

DNA samples should be shipped express or overnight at 4 ºC using ice packs or frozen using dry ice.
Please submit Sample Submission Form for Methylation Analysis with samples. If you are not with the University of Nebraska systems, please fill out the additional Payment Request Form. Thank you.

Chromatin Immunoprecipitation (ChIP)

For analysis involving Quantitative PCR (Real-Time QPCR) a minimum of 10 ng of clean reverse crosslinked DNA will be needed per PCR reaction. For High Throughput Sequencing Analysis (ChIP-Seq) a minimum of 80 ng is needed per treatment sample. DNA should be shipped express or overnight at 4 ºC using ice packs or frozen using dry ice. Please submit Sample Submission Form for ChIP Analysis with samples. If you are not with the University of Nebraska systems, please fill out the additional Payment Request Form. Thank you.

Gene Expression Analysis (QPCR)

Submitted RNA will be checked for degradation, genomic DNA contamination and purity (A260/280 ratio of 1.8 or above) using an Agilent Bioanalyzer. RNA should be treated with DNase I during the extraction procedure prior to submitting. RNA samples should be shipped overnight on dry ice. A minimum of 1 µg of RNA will be needed for one reverse transcription reaction, however, lower amounts can be utilized. Contact us with any special needs or applications. Please submit Sample Submission Form for QPCR Analysis with samples. If you are not with the University of Nebraska systems, please fill out the additional Payment Request Form. Thank you.