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to the DERC Website: PCR Net
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PCR Net |
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The Joslin Diabetes Center DERC PCR Core provides automated real-time PCR analysis of nucleic acids (e.g. quantitative RT-PCR, allelic discrimination, plus/minus). The PCR Core is part of the Genomics Core
Location: Room 616, Joslin Diabetes Center
Phone: 617-264-2795, internal ext. 4543
Research Technician: Josh Schroeder, Joshua.Schroeder@joslin.harvard.edu
Core Director: Mary Elizabeth Patti, Mary.Elizabeth.Patti@joslin.harvard.edu
Objective:
The main objective of the PCR Core is to facilitate research by DERC laboratories by providing equipment, expertise, and services in nucleic acid analysis that would be too specialized or costly for individual laboratories to provide independently. The PCR core currently provides automated real-time fluorescent PCR analysis of nucleic acids. Additionally, the core provides expertise in the design of oligonucleotides for various applications, including real-time RT-PCR.
Background:
The analysis and alteration of nucleic acid molecules continues to increase in importance for research in all areas of diabetes and endocrinology from the study of organismal physiology to cell biology and transcriptional regulation. Most DERC investigators need to quantify genes of interest either at the mRNA or DNA level; the ability to perform these measurements precisely using minimal starting material by fluorescent real-time PCR reduces waste of time and materials. The dedicated personnel and equipment required for this technique are not feasible for individual laboratories. The Joslin Diabetes Center PCR Core provides the expertise and equipment required for this important specialized technique.
Online Sign-up:
Please sign-up for slots online. E-mail Chris Botka for username and password if you have not used any of the Joslin Cores (not the same username and password as Joslin network).
PCR Sign-up Guidelines:
- On weekdays between 7:30 AM and 6:30 PM, please limit your sign-ups to 2 per day per machine.
- You may cancel your slot no later than 2 hours prior to start time if you do not plan on using it.
- Be advised, unused slots that are not cancelled will be charged.
- RT = Real-Time PCR (2 1/2 hour slot)
- AD = Allelic Discrimination (1/2 hour slot)
Equipment:
ABI PRISMŽ 7700 Sequence Detection System
ABI PRISMŽ 7000 Sequence Detection System
ABI PRISMŽ 7900 Sequence Detection System
Please see Applied Biosystems website for more information
MJ Research PTC-200 96-well Thermocycler
NanoDrop ND-1000
NanoDrop ND-3300
Please see NanoDrop website for more information
Please sign-up for NanoDrops. There is a sign-up sheet next to the machines. Thank you!
Services:
Please see RT-PCR resources at Applied Biosystems RT-PCR Support page
1. Assistance with setup of RT-PCR experiments (provide background information about RT-PCR, reagents needed, etc.).
2. Instruction with operation of SDS software and operation of ABI Sequence Detection Systems.
3. Assistance with design of primers and probes. Primer Express software is available to PCR Core users.
4. Assistance with analysis of results in SDS software and troubleshooting.
5. Primer database of validated target genes submitted by Joslin users.
Cost:
RT-PCR (2 1/2 hour slot): $25
Allelic discrimination (1/2 hour slot): $5
Optical plates/caps (if user is in short supply): $10 each
Other primer resources:
PrimerBank
Applied Biosystems
Quantitative PCR Primer Database (QPPD)
RTPrimerDB
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