BioMicroCenter:RTPCR: Difference between revisions

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== Traditional Real Time PCR ==


== [[BioMicroCenter:BioMark | Fluidigm BioMark]] ==
[[Image:BioMicroCenter_BioMarkChip.jpg|left]]


The Fluidigm BioMark system is a highly parallelized system allowing for running thousands of RT-PCR reactions on a single cartridge. Useful for genotyping, gene expression analysis and digital PCR, the BioMark cassette works using thousands of [http://www.fluidigm.com/glossary/gloss_nflex.htm NanoFlex(TM) valves] to fill thousands of individual reactions from a matrix of samples and primers. Standard Geneotyping / Gene Expression cassettes come in 48x48 and 96x96 format allowing either ~2,500 or ~10,000 reactions on a single cassette. RT-PCR in the BioMark system is compatible with TaqMan(TM), the Roche Universal priming kit, and SYBR Green technologies.


Using the BioMark system requires some additional processing over traditional RT-PCR. Using only 10nL in each reaction, ensuring that each reaction has sufficient template is a primary concern. To create a concentrated enough template, the BioMark system requires a pre-amplification where all of the primer pairs to be tested are mixed together with each sample and amplified by standard PCR. This increases the concentration of the specific product enough to ensure that sufficient template is added to each of the final reactions.
=== Light Cycler 480 II Real-time PCR Machines ===
 
{| align=left
* <B>[https://mit-ki.ilabsolutions.com/equipment/show/261686/?tab=schedule Schedule time on the Biomark]</B><BR><BR>
|- valign="top"
 
| <!-- BMC CORE TABLE -->
Use of the BioMark costs '''$150''' per use to help defray the service contract charge.
{| border="1" cellpadding="5" cellspacing="0" align=right
For training please email biomicro@mit.edu
  ! Instrument || LC480
 
  |-
== Traditional Real Time PCR ==
  | TYPE || WALKUP - MIT only
  |-
  | UNIT || Per Plate
  |-  
  | FORMAT || 96w/394w
  |-
  | SIGNUP || [https://mit.ilabsolutions.com/equipment/show/261680/?tab=schedule ilabs]
  |-
  | NEW USERS ||  New users should request training by emailing biomicro@mit.edu
  |-
  | DONATED BY || Prof Laurie Boyer, Prof Leonard Guarente, Dept of Biology
  |}
| <!-- DESCRIPTION -->
The [https://www.roche-applied-science.com/sis/rtpcr/htc/index.jsp LightCycler 480 Real-Time PCR System] is a fully integrated multiwell-plate based real-time PCR platform for highly accurate qualitative and quantitative detection of nucleic acids. The instrument handles both 96 and 384w plates, rapidly switching between blocks (protocol below). The LC480 can measure Ct and melt curves and can do protein melt curves as well. The instrument handles SYBR and Taqman probes. ROC standards are not required on the instrument, though it can detect them. <BR><BR>


[[Image:LC480.jpg|right]]
There is a charge per plate to cover the cost of the service contract.<BR><BR>


=== Light Cycler 480 II Real-time PCR Machines ===
96 well plates and SYBR Green can also be purchased from the BioMicro Center for use on the LightCycler480<BR><BR>
The [https://www.roche-applied-science.com/sis/rtpcr/htc/index.jsp LightCycler 480 Real-Time PCR System] is a fully integrated multiwell-plate based real-time PCR platform for highly accurate qualitative and quantitative detection of nucleic acids. Building on the benefits of Roche's capillary-based LightCycler® Systems, it goes one step further in offering enhanced throughput, compatibility with automation equipment and maximum flexibility regarding hard- and software. Providing novel ways to combine speed and accuracy without compromises, the LightCycler® 480 Real-Time PCR System meets the needs of a broad range of applications in research fields such as gene expression studies, discovery and analysis of genetic variation or array data validation.


There is a charge per plate to cover the cost of the service contract.  
'''Please Note:''' There are three LightCycler systems (Thing 1, Thing 2, and Thing 3) that have a sign up <B>maximum of four hours per day</b> during business hours (10AM to 5PM), all time slots are available the night before even if it exceeds the maximum.  


96 well plates and SYBR Green can also be supplied on demand for use on the LightCycler480


'''Please Note:''' There are three LightCycler systems (Thing 1, Thing 2, and Thing 3) that have a sign up maximum of four hours per day during business hours (10AM to 5PM), all time slots are available the night before even if it exceeds the maximum.
[https://mit-ki.ilabsolutions.com/equipment/show/261680/?tab=schedule Schedule Time on the LightCycler 480]
   
   
[[Image:Block Changing Protocol LightCycler480.txt]]
[[Image:Block Changing Protocol LightCycler480.txt]]
| <!-- IMAGE -->
[[Image:LC480.jpg|right]]
|}

Latest revision as of 20:37, 20 October 2018

HOME -- SEQUENCING -- LIBRARY PREP -- HIGH-THROUGHPUT -- COMPUTING -- OTHER TECHNOLOGY

Traditional Real Time PCR

Light Cycler 480 II Real-time PCR Machines

Instrument LC480
TYPE WALKUP - MIT only
UNIT Per Plate
FORMAT 96w/394w
SIGNUP ilabs
NEW USERS New users should request training by emailing biomicro@mit.edu
DONATED BY Prof Laurie Boyer, Prof Leonard Guarente, Dept of Biology

The LightCycler 480 Real-Time PCR System is a fully integrated multiwell-plate based real-time PCR platform for highly accurate qualitative and quantitative detection of nucleic acids. The instrument handles both 96 and 384w plates, rapidly switching between blocks (protocol below). The LC480 can measure Ct and melt curves and can do protein melt curves as well. The instrument handles SYBR and Taqman probes. ROC standards are not required on the instrument, though it can detect them.

There is a charge per plate to cover the cost of the service contract.

96 well plates and SYBR Green can also be purchased from the BioMicro Center for use on the LightCycler480

Please Note: There are three LightCycler systems (Thing 1, Thing 2, and Thing 3) that have a sign up maximum of four hours per day during business hours (10AM to 5PM), all time slots are available the night before even if it exceeds the maximum.


File:Block Changing Protocol LightCycler480.txt

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