Services Offered
Nucleic Acid Purification
Qiagen Qiacube (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Nucleic acid isolation and purification | Total RNA; miRNA; DNA; All prep (all nucleic acid) 2-12 samples per run | Fully automated; DNase over column; Flexible to handle cells and tissues | Efficiency of columns limited | Specialized methods needed for miRNA. Appointment and training required |
Nucleic Acid Quantification and Quality Control
Agilent Bioanalyzer (Contact: Zoe Brookover)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Quality Control | Profiles for total RNA; enriched mRNA; enriched small RNA; DNA fragment up to ~10kb; | Measure integrity and quantity of DNA and RNA; samples sizes down to ~100pg | Quantification not as accurate as other methods; salt skews results | Samples must be in nuclease free water. |
Nanodrop Spectrophotometer (Contact: MRC @ 901.448.6191)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Nucleic acids quantification and quality control | Absorbance for nucleic acids; calculating concentrations and purity; A260/A280 ratios; and A260/A230 ratios | Able to see contamination of organics and proteins; requires only 1µl of sample | Loses accuracy at concentrations below 20ng/µl and above 1500ng/µl. Concentrations affected by purity. | Nucleic acids only |
Qubit (Contact: Zoe Brookover)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Quantification of nucleic acids | Florescence detection for concentration of nucleic acids | Organic contamination not usually a problem; high sensitivity | Does not allow determination of sample purity. | Needs 1µl of sample |
Spectra Max M2e Plate Reader (Contact: Zoe Brookover)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Quantification of nucleic acids and proteins | Absorbance; fluorescence; luminescence for DNA; RNA; and protein assays | Reads plates or cuvette with a wide range of wave lengths | User must provide plates or cuvette | Works with any flat bottom 96 well plate. Prior training necessary. |
Gene Expression and Genetic Variation (High Throughput)
Life Technologies Ion Torrent PGM (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Next generation sequencing; genetic variation and discovery | ChIP seq; miRNA seq; targeted seq; microbiome; 1-96 bar-coded samples for DNA and 1-48 bar-coded samples for RNA | Good for discovery experiments; lower cost with high throughput; very flexible | Restricted amount of coverage | Arrange for bioinformatics consultation before run. Prior consultation with MRC required to prepare a quote. |
Life Technologies Proton (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Next generation sequencing; genetic variation and discovery | RNA-seq; ChIP seq; miRNA seq; whole genome seq; exome sequencing; 1-96 bar-coded samples for DNA and 1-48 bar-coded samples for RNA; scalable coverage |
De novo sequencing; differential gene expression and detection of splicing variants; SNP detection | Low abundant transcripts will require deeper coverage for detection; for RNA-seq, must have a reference library; fewer downstream workflows available than for Illumina NGS data | Arrange for bioinformatics consultation before run. Prior consultation with MRC required to prepare a quote. |
Illumina NextSeq2000 (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
| Next generation sequencing; genetic variation and discovery | chIP-seq; miRNA-seq; targeted sequencing; whole genome sequencing; exome sequencing; 1-384 bar-coded samples; scalable coverage | De novo sequencing; differential gene expression and detection of splicing variants; SNP detection; many downstream raw data analysis workflows available | Low abundant transcripts will require deeper coverage for detection; for RNA-seq, must have a reference library | Arrange for bioinformatics consultation before run. Prior consultation with MRC required to prepare a quote. |
Affymetrix Microarray (Contact: Zoe Brookover)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
| Gene expression and genetic variation | Differential gene expression; alternative splicing; genotyping; cytogenetics; miRNA; 1 sample/array | Whole transcript (no 3' bias); high correlation values; gene level and exon level data; excellent for miRNA detection; data easy to analyze | Restricted to probes tiled on the array; hybridizaton based application; less reliable for low expression | Requires validation. Prior consultation with MRC required. |
Real Time PCR and Genetic Variation
Roche LC480 (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Real-time PCR | Gene expression; genotyping; HRM; Taqman UPL library®; Sybr Green 96- and 384-well formats |
Highly sensitive; reproducible and flexible UPL Library available for human; mouse and rat | Sybr green assays prone to errors from primer dimers and nonspecific amplification | Reference gene assays available. There is limited availability of UPL probes. Contact the MRC for more information. |
Fluidigm Biomark (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Real-time PCR | qRT-PCR; digital PCR; genotyping Taqman UPL library®; Sybr green Various high throughput formats | Cost effective for high throughput; low sample input requirements; highly sensitive | Defined formats determine number of samples and genes | Samples must be pre-amplified |
Other Services
Eppendorf epMotion 5075 (Contact: Zoe Brookover)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Liquid handling robot | Liquid transfer to 384 well plates | Able to quickly set up 384 well plates for Roche LC480 | Must dispense either single or 8 channels | Prior appointment necessary |
Zeiss Axiophot Photomicroscope (Contact: Bill Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Fluorescence microscopy | Fluorescence detection assays; photo documentation available | Progres® software able to take color pictures of fluorescence detection assays | Limited resolution. Dyes limited to Red (PE's); Green (Fitc); DAPI or dyes with similar spectra | Prior appointment and training necessary |
Services by External Vendors
Azenta (Contact: William Taylor)
| Application | Capabilities | Advantages | Requirements | Pricing of Pre-Mixed Samples |
|---|---|---|---|---|
| Sanger DNA sequencing | plasmid DNA and PCR products | Next-day results, if received in the MRC dropbox by 2:30pm CST. Quality control reports provided with data, free technical support and a re-sequencing policy. |
please refer to the "Sample Submission Guidelines"
When signing into your online, you must choose the MRC as the bill to location: MRC, Rene' Smira, 152 CRB, 19 S. Manassas St, Memphis TN 38163, 901-448-2491 and you must reference this code in the PO field "MRC42473". Users will be billed list price for Azenta services if these guidelines are not followed.
|
For current price information, please contact Tiffany Seagroves, or contact Azenta directly at 908-222-0711 and request
the negotiated pricing for UTHSC. |
Transnetyx Genotyping (Contact: William Taylor)
| Application | Capabilities | Advantages | Disadvantages | Recommendations |
|---|---|---|---|---|
| Genotyping | Knockouts; transgenic mutations; SNP detection using real-time PCR | Fully automated process; mail in mouse tail or ear | NA | Check with company for acceptable tissues to send; drop off kiosk located at MRC |
Molecular Resource Center External Sanger Sequencing Submission Policy Change
Samples submitted at the MRC for Sanger sequencing by Azenta (formerly GeneWiz) must be logged into the iLab portal by the laboratory personnel submitting the samples, in addition to completion of the external submission form on the Azenta portal. Both the iLab and Azenta paperwork must be downloaded, printed and submitted along with your samples for sequencing, or they will not be sent out.