The quality of your data will be directly proportional to the quality of your starting material. Modern capillary sequencing, Next-Gen sequencing, quantitative PCR, droplet PCR, and the whole host of single cell applications available have more stringent requirements for high quality nucleic acids than traditional techniques.
To help our investigators prepare high quality nucleic acids, we routinely test different kits for their ability to REPRODUCIBLY generate high quality sample. We have sequenced and amplified literally HUNDREDS of THOUSANDS of samples and currently recommend the following methods and kits.
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Promega Maxwell 16 Automated Nucleic Acid Extraction
Maxwell provides an automated process of extracting high quality DNA and RNA from
a variety of sample types. Using prefilled cartridges, the Maxwell delivers fast,
reliable and consistent DNA and RNA, along with the flexibility to process from 1
to 16 samples per run.Several Promega Maxwell 16 kits are available based onDNA or RNA as well as downstream application, sample source (tissue, cells, etc.), and elution volume (SEV or LEV).
A member of your core will set up appropriate hardware for the kit used. User fee applied.
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Recommended Kits for Manual DNA Isolation/ Purification
For Most Downstream Applications
- PureLink DNA Mini and Midi Kits: Some of the longest sequencing reads we've obtained used these kits for the DNA isolation
ChargeSwitch Kits: One of the Director's personal favorites for ease and quality of the DNA isolation
QIAGEN DNeasy Blood and Tissue Kits: Routinely generate excellent quality DNA
Be careful not to have any leftover EtOH in your prep as this is detrimental to downstream applications.
For Next-Gen Sequencing of Microbiome Samples
- QIAamp PowerFecal Pro DNA Kit: Includes a bead-beating step that is crucial to ensure complete lysis of ALL bacteria, preventing sample biasing towards Gram negative organisms.
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Recommended Kits for Manual RNA Isolation
For Most Downstream Applications
- QIAGEN RNeasy Kits: Includes an on-column DNase digestion using RNase-free DNase set. Available in micro to maxi prep size and 96 well formats
- Ambion RNAqueous Mini and Midi Kits : Phenol-free RNA isolation. Phenol is a common contaminant that can alter RNA concentration measurements and inhibit PCR
For High Throughput
- Promega Automated SV96 Total RNA Isolation System: Can be used on the Beckman Biomek 2000 robot, present in your core. This kit is designed to isolate total RNA from cultured cells but we have used successfully isolated RNA from animal tissue (rat brain)