BioMicroCenter:RTPCR - Revision history

Stuart S. Levine: /* Traditional Real Time PCR */

Stuart S. Levine — Wed, 06 Feb 2013 03:39:45 GMT

Traditional Real Time PCR

←Older revision		Revision as of 03:39, 6 February 2013
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	== Traditional Real Time PCR ==		== Traditional Real Time PCR ==
-
-	~~[[Image:Opticon2_system.gif\|right]]~~
-	~~===MJ Research Real Time PCR Machine===~~
-
-	The MJ Research PCR Machine enables multicolor capability allowing detection of SYBR® Green I or FAM in the first channel, and a range of fluorophores in the second channel — including TET, HEX, VIC, and TAMRA — for a multitude of applications such as RT-qPCR and allelic discrimination. The DNA Engine® thermal cycler offers precision thermal control and a temperature gradient feature permits simultaneous incubation at 12 different temperatures to optimize reactions in a single run. The real-time results allow for plotting of signal intensity vs. cycle number and graphically monitoring the thermal profile during the run. The extraordinary sensitivity permits reliable detection of one initial template copy, while delivering a linear range of up to ten orders of magnitude in starting copy number (with appropriate chemistry). Users are currently charged '''$10''' per plate for the use of this machine. <BR>
-	* <B>[[BioMicroCenter:MJO_CAL\|Schedule Time on the MJ Opticon]]</B><BR><BR>

	[[Image:LC480.jpg\|right]]		[[Image:LC480.jpg\|right]]
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	[[Image:Block Changing Protocol LightCycler480.txt]]		[[Image:Block Changing Protocol LightCycler480.txt]]
		+
		+
		+	[[Image:Opticon2_system.gif\|right]]
		+	===MJ Research Real Time PCR Machine===
		+
		+	''' THE MJ IS CURRENTLY RETIRED WHILE THE WAFERGEN SYSTEM IS IN PLACE '''
		+	The MJ Research PCR Machine enables multicolor capability allowing detection of SYBR® Green I or FAM in the first channel, and a range of fluorophores in the second channel — including TET, HEX, VIC, and TAMRA — for a multitude of applications such as RT-qPCR and allelic discrimination. The DNA Engine® thermal cycler offers precision thermal control and a temperature gradient feature permits simultaneous incubation at 12 different temperatures to optimize reactions in a single run. The real-time results allow for plotting of signal intensity vs. cycle number and graphically monitoring the thermal profile during the run. The extraordinary sensitivity permits reliable detection of one initial template copy, while delivering a linear range of up to ten orders of magnitude in starting copy number (with appropriate chemistry). Users are currently charged '''$10''' per plate for the use of this machine. <BR>
		+	* <B>[[BioMicroCenter:MJO_CAL\|Schedule Time on the MJ Opticon]]</B><BR><BR>

Stuart S. Levine: /* Wafergen Smartchip */

Stuart S. Levine — Wed, 06 Feb 2013 03:38:30 GMT

Wafergen Smartchip

←Older revision		Revision as of 03:38, 6 February 2013
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	== [[BioMicroCenter:Wafergen \| Wafergen Smartchip]] ==		== [[BioMicroCenter:Wafergen \| Wafergen Smartchip]] ==
-	[[Image:~~BioMicroCenter_Smartchip~~.jpg\|~~left~~]]	+	[[Image:BioMicroCenter-Smartchip.jpg\|300px\|right]]

	The Wafergen Smartchip system is a highly parallelized system allowing for running thousands of RT-PCR reactions on a single cartridge. Useful for genotyping and gene expression, the Smartchip works using very small reaction volumes (100nL) pipetted with a specialized automated liquid handler. The 72x72 chip can be filled in a broad variety of patterns. RT-PCR in the Wafergen system is compatible with TaqMan(TM), the Roche Universal priming kit, and SYBR Green technologies.		The Wafergen Smartchip system is a highly parallelized system allowing for running thousands of RT-PCR reactions on a single cartridge. Useful for genotyping and gene expression, the Smartchip works using very small reaction volumes (100nL) pipetted with a specialized automated liquid handler. The 72x72 chip can be filled in a broad variety of patterns. RT-PCR in the Wafergen system is compatible with TaqMan(TM), the Roche Universal priming kit, and SYBR Green technologies.
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	* <B>[[BioMicroCenter:Wafer_CAL\| Schedule time on the Wafergen]]</B><BR><BR>		* <B>[[BioMicroCenter:Wafer_CAL\| Schedule time on the Wafergen]]</B><BR><BR>
-
-

	== [[BioMicroCenter:BioMark \| Fluidigm BioMark]] ==		== [[BioMicroCenter:BioMark \| Fluidigm BioMark]] ==

Stuart S. Levine at 03:09, 6 February 2013

Stuart S. Levine — Wed, 06 Feb 2013 03:09:31 GMT

←Older revision		Revision as of 03:09, 6 February 2013
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	{{BioMicroCenter}}		{{BioMicroCenter}}
		+
		+	== [[BioMicroCenter:Wafergen \| Wafergen Smartchip]] ==
		+	[[Image:BioMicroCenter_Smartchip.jpg\|left]]
		+
		+	The Wafergen Smartchip system is a highly parallelized system allowing for running thousands of RT-PCR reactions on a single cartridge. Useful for genotyping and gene expression, the Smartchip works using very small reaction volumes (100nL) pipetted with a specialized automated liquid handler. The 72x72 chip can be filled in a broad variety of patterns. RT-PCR in the Wafergen system is compatible with TaqMan(TM), the Roche Universal priming kit, and SYBR Green technologies.
		+
		+	The Wafergen Smartchip system is currently on loan within the BioMicro Center. Future availability will be based on usage level.
		+
		+	* <B>[[BioMicroCenter:Wafer_CAL\| Schedule time on the Wafergen]]</B><BR><BR>
		+
		+

	== [[BioMicroCenter:BioMark \| Fluidigm BioMark]] ==		== [[BioMicroCenter:BioMark \| Fluidigm BioMark]] ==

Kayla L. Menard: /* Light Cycler 480 II Real-time PCR Machines */

Kayla L. Menard — Thu, 18 Oct 2012 18:56:12 GMT

Light Cycler 480 II Real-time PCR Machines

←Older revision		Revision as of 18:56, 18 October 2012
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	* Password: DNARNAprotein		* Password: DNARNAprotein

-	[[Image:Block Changing Protocol ~~for Roche~~ LightCycler480.~~docx~~]]	+	[[Image:Block Changing Protocol LightCycler480.txt]]

Kayla L. Menard: /* Light Cycler 480 II Real-time PCR Machines */

Kayla L. Menard — Tue, 16 Oct 2012 17:52:09 GMT

Light Cycler 480 II Real-time PCR Machines

←Older revision		Revision as of 17:52, 16 October 2012
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	* Username: Building68		* Username: Building68
	* Password: DNARNAprotein		* Password: DNARNAprotein
		+
		+	[[Image:Block Changing Protocol for Roche LightCycler480.docx]]

Shmulik Motola at 16:23, 6 March 2012

Shmulik Motola — Tue, 06 Mar 2012 16:23:46 GMT

←Older revision		Revision as of 16:23, 6 March 2012
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	* Username: Building68		* Username: Building68
	* Password: DNARNAprotein		* Password: DNARNAprotein
-
-	~~== PCR ==~~
-
-	~~[[Image:Primus_multiblock.jpg\|right]]~~
-	~~===MWG Thermal Cyclers===~~
-	~~The Primus Multiblock system provides comprehensive state-of-the-art PCR technology.~~
-	~~They are ideal instruments for high throughput PCR and sequencing. The cyclers allow for~~
-	~~exchangeable blocks (384/96 well format), motorized lid, HTR Peltier technology for~~
-	~~fastest heating / cooling rates and optimal results. Patented HPL (High Pressure Lid)~~
-	~~technology for 100% evaporation prevention.~~
-
-	The MWG PCR machines are available for non-BMC labs at a $10 charge. If you are interested in the machines you must coordinate use with BioMicro Center staff as the machines are used extensively in many protocols run by the staff.

Stuart S. Levine: /* Fluidigm BioMark */

Stuart S. Levine — Tue, 31 Jan 2012 03:05:49 GMT

Fluidigm BioMark

←Older revision		Revision as of 03:05, 31 January 2012
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	* <B>[[BioMicroCenter:Fluid_CAL\| Schedule time on the Biomark]]</B><BR><BR>		* <B>[[BioMicroCenter:Fluid_CAL\| Schedule time on the Biomark]]</B><BR><BR>

-	Use of the BioMark costs '''$~~100~~''' per use to help defray the service contract charge.	+	Use of the BioMark costs '''$150''' per use to help defray the service contract charge.
	For training please email biomicro@mit.edu		For training please email biomicro@mit.edu

Justin Elliott: /* Light Cycler 480 II Real-time PCR Machines */

Justin Elliott — Mon, 13 Jun 2011 14:04:39 GMT

Light Cycler 480 II Real-time PCR Machines

←Older revision		Revision as of 14:04, 13 June 2011
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	96 well plates ($6 per plate) and SYBR Green ($40 per mL) can also be supplied on demand for use on the LightCycler480		96 well plates ($6 per plate) and SYBR Green ($40 per mL) can also be supplied on demand for use on the LightCycler480

-	'''Please Note:''' There are ~~two~~ LightCycler systems (Thing 1 ~~and~~ Thing 2) 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. New users who wished to be trained must be [[BioMicroCenter:CoreDeps\|Core Lab members]].	+	'''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. New users who wished to be trained must be [[BioMicroCenter:CoreDeps\|Core Lab members]].

	[http://www.google.com/calendar Schedule Time on the LightCycler 480]		[http://www.google.com/calendar Schedule Time on the LightCycler 480]

Ryan Sinapius: /* Fluidigm BioMark */

Ryan Sinapius — Fri, 21 Jan 2011 16:07:02 GMT

Fluidigm BioMark

←Older revision		Revision as of 16:07, 21 January 2011
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	* <B>[[BioMicroCenter:Fluid_CAL\| Schedule time on the Biomark]]</B><BR><BR>		* <B>[[BioMicroCenter:Fluid_CAL\| Schedule time on the Biomark]]</B><BR><BR>

-	Use of the BioMark costs '''$100''' per ~~cassette~~ to help defray the service contract charge.	+	Use of the BioMark costs '''$100''' per use to help defray the service contract charge.
	For training please email biomicro@mit.edu		For training please email biomicro@mit.edu

Stuart S. Levine at 16:11, 22 September 2010

Stuart S. Levine — Wed, 22 Sep 2010 16:11:14 GMT

←Older revision		Revision as of 16:11, 22 September 2010
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	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.		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.

-	* [[BioMicroCenter:Fluid_CAL\|~~''' Sign Up for~~ time on the Biomark~~'''~~]]	+	* <B>[[BioMicroCenter:Fluid_CAL\| Schedule time on the Biomark]]</B><BR><BR>

	Use of the BioMark costs '''$100''' per cassette to help defray the service contract charge.		Use of the BioMark costs '''$100''' per cassette to help defray the service contract charge.