BioMicroCenter:PacBio - Revision history

Asoberan: Redirect to consolidated BioMicroCenter:LongRead page

2026-04-21T01:49:35Z

Redirect to consolidated BioMicroCenter:LongRead page

← Older revision		Revision as of 01:49, 21 April 2026
Line 1:		Line 1:
	~~{{BioMicroCenter}}~~		#REDIRECT [[BioMicroCenter:LongRead]]

	~~== PACBIO REVIO==~~
	{\|
	~~\|- style="vertical-align: top;"~~
	~~\|style="width: 400px;"\|~~
	~~{\| class="wikitable" border=1~~
	!
	~~!REVIO SEQUENCING~~
	\|-
	~~\|CHEMISTRY <BR> FLOWCELL~~
	~~\| [https://www.pacb.com/revio/ PacBio Revio] <BR> 10 M wells loaded by poissonian distribution~~
	\|-
	~~\|INPUT \|\| Completed SMRTbell Libraries~~
	* 12uL
	* >1nM
	~~''1nM ~ 0.65ng/ ul/ kb insert''~~
	\|-
	~~\|INCLUDED SERVICES \|\|~~
	* Sample QC on [[BioMicroCenter:QC#AATI_FEMTO_PULSE\|FemtoPulse]]
	* Sequencing
	* Primary analysis
	* Data storage for 1y
	\|-
	~~\|ADDITIONAL SERVICES \|\|~~
	*[[BioMicroCenter:PacBio_Library_Preparation\|PacBio Library Prep]]
	*[[BioMicroCenter:PippinPrep\|Pippin Prep]]
	\|-
	~~\|PRICING \|\| [[BioMicroCenter:Pricing~~#~~PACBIO_SEQUENCING\|LINK]]~~
	\|-
	~~\|SUBMISSION \|\| MIT - [https://mit.ilabsolutions.com/service_item/new/3381?spt_id=9912 ilabs] <BR> External -~~ [[BioMicroCenter:~~Forms\|form]]~~
	\|-
	~~\|PROVIDED WITH \|\|~~
	~~Local academic cores~~
	\|}
	\|
	~~[[Image:sequel.jpeg\|right]]~~
	\|}

	~~== [http://www.pacb.com/products/smrt-technology/ HOW SINGLE MOLECULE SEQUENCING WORKS ]==~~
	The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. Ideally, a PacBio library with a single polymerase is loaded into each well. When a fluorescent nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>
	The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>

	~~[[IMAGE:CCS-demo.jpg\|thumb\|left\|400px\|Diagram of CCS analysis]]~~

	Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 24 hr movie. Rare lengths of 175 kbp have been seen, but it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.

	~~<!-- Commented out related to Sequel~~
	~~== EXPECTED RESULTS ==~~
	~~TYPICAL RESULTS FROM SMRTCELLS:~~
	* 20kb fragments – 50,000 active sites – 400Mbp sequence
	* 2kb fragments – 80,000 active sites – 500Mbp sequence

	~~Typical Read Lengths~~
	~~{\|border=1 align="left"~~
	\|-
	~~!Genomic!!Amplicon (<2 kb)~~
	\|-
	\|[[IMAGE:PacBioSequelTypicalv2.jpg\|thumb\|left\|400px\|Genomic DNA prepared with Template Kit 1.0 and BluePippin on Sequel 2.1 Chemistry]]\|\|[[IMAGE:PacBioSequelTypicalv2-amp.jpg\|thumb\|left\|400px\|PCR Amplicon prepared with Express Template Kit 2.0 on Sequel 3.0 Chemistry]]
	\|-
	~~\|Longest Inserts of around 60 kbp in CLR\|\|Shorter Inserts mean higher Quality Scores in CCS~~
	\|}
	~~<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br> -->~~

	~~== APPLICATIONS ==~~
	~~There are several applications for which PacBio sequencing are well suited (and many it is not!). The major ones are below:~~
	~~=== [http://www.pacb.com/applications/denovo/ DE NOVO ASSEMBLY / LARGE SCALE MAPPING~~] ~~===~~
	Assembly is the biggest strength of PacBio. While the individual base error rate is quite high (~14%), the errors are random and so resequencing rapidly lowers the error rate. Using only PacBio reads, 60x coverage can give very good assembly – typically completing genomes in 1 shot . A 4 Mbp genome can be done on a single SMRT cell. <BR><BR>
	A second strategy is to use PacBio reads to supplement short reads and join scaffolds. Here, standard short reads make up the bulk of the assembly but the separate scaffolds can be spanned by PacBio reads. 7x reads has been the convention for this type of sequencing. 400 Mbp of genomic DNA can be done on a single SMRT cell. Indexing cannot be done easily with PacBio samples - particularly long reads - and so the 'quanta' for SMRTcells should be considered as up to three genomes per SMRTcell.

	~~=== [http://www.pacb.com/applications/target/index.html AMPLICON / RESEQUENCING] ===~~
	~~PacBio reads can also be used to looks for variants in specific regions. The long reads allow for better detection of large rearrangements and understanding repetitive regions of the genome. <BR><BR>~~
	The use of long reads in assembly can also establish phasing of mutations. Short reads will typically not be able to span multiple mutations on a single read. PacBio reads are long enough to enable detection of multiple mutations as coming from the same strand. <BR><BR>
	~~Indexing is possible with resequencing amplicons but indexes should be added during PCR and not during library preparation.~~

	~~=== [https://www.pacb.com/applications/rna-sequencing/ WHOLE TRANSCRIPT] ===~~
	Unlike short read RNAseq, which requires fragmenting of the RNA, full length cDNAs can be sequenced on the PacBio Sequel. This allows direct detection of different splice isoforms. The low number of reads makes using this as a counting method challenging.

	~~=== [http://www.pacb.com/applications/base_modification/index.html BACTERIAL BASE MODIFICATION~~] ~~===~~
	One aspect that makes PacBio sequencing unique is its ability to recognize chemical modifications of bases. Because the instrument works by measuring the time a nucleotide is in the active site, it has an axis of “time”. Basepairing modified bases has different kinetics which allows the sequencer to provide a likelihood score that any given base has been modified. Methylcytosine, Hydroxymethylcytosine, 6-methyl adenosine and many others have been detected utilizing this kinetics mode.

Hilo1: /* HOW SINGLE MOLECULE SEQUENCING WORKS */

2025-06-09T17:52:11Z

HOW SINGLE MOLECULE SEQUENCING WORKS

← Older revision		Revision as of 17:52, 9 June 2025
Line 41:		Line 41:
	The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. Ideally, a PacBio library with a single polymerase is loaded into each well. When a fluorescent nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>		The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. Ideally, a PacBio library with a single polymerase is loaded into each well. When a fluorescent nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>
	The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>		The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>

			[[IMAGE:CCS-demo.jpg\|thumb\|left\|400px\|Diagram of CCS analysis]]

	Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 24 hr movie. Rare lengths of 175 kbp have been seen, but it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.		Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 24 hr movie. Rare lengths of 175 kbp have been seen, but it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.

Hilo1: /* BACTERIAL BASE MODIFICATION */

2025-06-09T17:36:57Z

BACTERIAL BASE MODIFICATION

← Older revision		Revision as of 17:36, 9 June 2025
Line 75:		Line 75:

	=== [http://www.pacb.com/applications/base_modification/index.html BACTERIAL BASE MODIFICATION] ===		=== [http://www.pacb.com/applications/base_modification/index.html BACTERIAL BASE MODIFICATION] ===
	One aspect that makes PacBio sequencing unique is its ability to recognize chemical modifications of bases. Because the instrument works by measuring the time a nucleotide is in the active site, it has an axis of “time”. Basepairing modified bases has different kinetics which allows the sequencer to provide a likelihood score that any given base has been modified. Methylcytosine, Hydroxymethylcytosine, 6-methyl adenosine and many others have been detected ~~with the RSII~~.		One aspect that makes PacBio sequencing unique is its ability to recognize chemical modifications of bases. Because the instrument works by measuring the time a nucleotide is in the active site, it has an axis of “time”. Basepairing modified bases has different kinetics which allows the sequencer to provide a likelihood score that any given base has been modified. Methylcytosine, Hydroxymethylcytosine, 6-methyl adenosine and many others have been detected utilizing this kinetics mode.

Hilo1: /* HOW SINGLE MOLECULE SEQUENCING WORKS */

2025-06-09T17:36:02Z

HOW SINGLE MOLECULE SEQUENCING WORKS

← Older revision		Revision as of 17:36, 9 June 2025
Line 39:		Line 39:

	== [http://www.pacb.com/products/smrt-technology/ HOW SINGLE MOLECULE SEQUENCING WORKS ]==		== [http://www.pacb.com/products/smrt-technology/ HOW SINGLE MOLECULE SEQUENCING WORKS ]==
	The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. A PacBio library with a single polymerase is loaded into each well. When a nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>		The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. Ideally, a PacBio library with a single polymerase is loaded into each well. When a fluorescent nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>
	The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>		The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>
	Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 24 hr movie. Rare lengths of 175 kbp have been seen, but it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.		Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 24 hr movie. Rare lengths of 175 kbp have been seen, but it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.

Hilo1: /* HOW SINGLE MOLECULE SEQUENCING WORKS */

2025-04-30T18:00:57Z

HOW SINGLE MOLECULE SEQUENCING WORKS

← Older revision		Revision as of 18:00, 30 April 2025
Line 41:		Line 41:
	The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. A PacBio library with a single polymerase is loaded into each well. When a nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>		The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. A PacBio library with a single polymerase is loaded into each well. When a nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>
	The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>		The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>
	Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 20 hr movie. ~~A mode called Continuous Long Read Sequencing simply sequences a subread sequenced over the length of the run, either 10 or 20 hours. In CLR mode, rare~~ lengths of 175 kbp have been seen, but ~~in the Center,~~ it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.		Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 24 hr movie. Rare lengths of 175 kbp have been seen, but it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.

	<!-- Commented out related to Sequel		<!-- Commented out related to Sequel

Hilo1 at 13:42, 25 April 2025

2025-04-25T13:42:23Z

← Older revision		Revision as of 13:42, 25 April 2025
Line 43:		Line 43:
	Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 20 hr movie. A mode called Continuous Long Read Sequencing simply sequences a subread sequenced over the length of the run, either 10 or 20 hours. In CLR mode, rare lengths of 175 kbp have been seen, but in the Center, it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.		Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 20 hr movie. A mode called Continuous Long Read Sequencing simply sequences a subread sequenced over the length of the run, either 10 or 20 hours. In CLR mode, rare lengths of 175 kbp have been seen, but in the Center, it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.

			<!-- Commented out related to Sequel
	== EXPECTED RESULTS ==		== EXPECTED RESULTS ==
	TYPICAL RESULTS FROM SMRTCELLS:		TYPICAL RESULTS FROM SMRTCELLS:
Line 57:		Line 58:
	\|Longest Inserts of around 60 kbp in CLR\|\|Shorter Inserts mean higher Quality Scores in CCS		\|Longest Inserts of around 60 kbp in CLR\|\|Shorter Inserts mean higher Quality Scores in CCS
	\|}		\|}
	<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>		<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br> -->

	== APPLICATIONS ==		== APPLICATIONS ==

Hilo1: /* DE NOVO ASSEMBLY / LARGE SCALE MAPPING */

2025-04-23T18:26:53Z

DE NOVO ASSEMBLY / LARGE SCALE MAPPING

← Older revision		Revision as of 18:26, 23 April 2025
Line 63:		Line 63:
	=== [http://www.pacb.com/applications/denovo/ DE NOVO ASSEMBLY / LARGE SCALE MAPPING] ===		=== [http://www.pacb.com/applications/denovo/ DE NOVO ASSEMBLY / LARGE SCALE MAPPING] ===
	Assembly is the biggest strength of PacBio. While the individual base error rate is quite high (~14%), the errors are random and so resequencing rapidly lowers the error rate. Using only PacBio reads, 60x coverage can give very good assembly – typically completing genomes in 1 shot . A 4 Mbp genome can be done on a single SMRT cell. <BR><BR>		Assembly is the biggest strength of PacBio. While the individual base error rate is quite high (~14%), the errors are random and so resequencing rapidly lowers the error rate. Using only PacBio reads, 60x coverage can give very good assembly – typically completing genomes in 1 shot . A 4 Mbp genome can be done on a single SMRT cell. <BR><BR>
	A second strategy is to use PacBio reads to supplement ~~Illumina~~ reads to join scaffolds. Here, standard ~~MiSeq~~ reads make up the bulk of the assembly but the separate scaffolds ~~are~~ spanned by PacBio reads. 7x reads has been the convention for this type of sequencing. 40 Mbp of genomic DNA can be done on a single SMRT cell. Indexing cannot be done easily with PacBio samples - particularly long reads - and so the 'quanta' for SMRTcells should be considered as ~~1 genome~~ per SMRTcell.		A second strategy is to use PacBio reads to supplement short reads and join scaffolds. Here, standard short reads make up the bulk of the assembly but the separate scaffolds can be spanned by PacBio reads. 7x reads has been the convention for this type of sequencing. 400 Mbp of genomic DNA can be done on a single SMRT cell. Indexing cannot be done easily with PacBio samples - particularly long reads - and so the 'quanta' for SMRTcells should be considered as up to three genomes per SMRTcell.

	=== [http://www.pacb.com/applications/target/index.html AMPLICON / RESEQUENCING] ===		=== [http://www.pacb.com/applications/target/index.html AMPLICON / RESEQUENCING] ===

Hilo1: /* WHOLE TRANSCRIPT */

2025-04-23T18:25:53Z

WHOLE TRANSCRIPT

← Older revision		Revision as of 18:25, 23 April 2025
Line 71:		Line 71:

	=== [https://www.pacb.com/applications/rna-sequencing/ WHOLE TRANSCRIPT] ===		=== [https://www.pacb.com/applications/rna-sequencing/ WHOLE TRANSCRIPT] ===
	Unlike ~~Illumina~~ RNAseq, which requires fragmenting of the RNA, full length cDNAs can be sequenced on the PacBio Sequel. This allows direct detection of different splice isoforms. The low number of reads makes using this as a counting method challenging.		Unlike short read RNAseq, which requires fragmenting of the RNA, full length cDNAs can be sequenced on the PacBio Sequel. This allows direct detection of different splice isoforms. The low number of reads makes using this as a counting method challenging.

	=== [http://www.pacb.com/applications/base_modification/index.html BACTERIAL BASE MODIFICATION] ===		=== [http://www.pacb.com/applications/base_modification/index.html BACTERIAL BASE MODIFICATION] ===
	One aspect that makes PacBio sequencing unique is its ability to recognize chemical modifications of bases. Because the instrument works by measuring the time a nucleotide is in the active site, it has an axis of “time”. Basepairing modified bases has different kinetics which allows the sequencer to provide a likelihood score that any given base has been modified. Methylcytosine, Hydroxymethylcytosine, 6-methyl adenosine and many others have been detected with the RSII.		One aspect that makes PacBio sequencing unique is its ability to recognize chemical modifications of bases. Because the instrument works by measuring the time a nucleotide is in the active site, it has an axis of “time”. Basepairing modified bases has different kinetics which allows the sequencer to provide a likelihood score that any given base has been modified. Methylcytosine, Hydroxymethylcytosine, 6-methyl adenosine and many others have been detected with the RSII.

Hilo1: /* AMPLICON / RESEQUENCING */

2025-04-23T18:25:41Z

AMPLICON / RESEQUENCING

← Older revision		Revision as of 18:25, 23 April 2025
Line 67:		Line 67:
	=== [http://www.pacb.com/applications/target/index.html AMPLICON / RESEQUENCING] ===		=== [http://www.pacb.com/applications/target/index.html AMPLICON / RESEQUENCING] ===
	PacBio reads can also be used to looks for variants in specific regions. The long reads allow for better detection of large rearrangements and understanding repetitive regions of the genome. <BR><BR>		PacBio reads can also be used to looks for variants in specific regions. The long reads allow for better detection of large rearrangements and understanding repetitive regions of the genome. <BR><BR>
	The use of long reads in assembly can also establish phasing of mutations. Short ~~Illumina~~ reads will typically not be able to span multiple mutations on a single read. PacBio reads are long enough to enable detection of multiple mutations as coming from the same strand. <BR><BR>		The use of long reads in assembly can also establish phasing of mutations. Short reads will typically not be able to span multiple mutations on a single read. PacBio reads are long enough to enable detection of multiple mutations as coming from the same strand. <BR><BR>
	Indexing is possible with resequencing amplicons but indexes should be added during PCR and not during library preparation.		Indexing is possible with resequencing amplicons but indexes should be added during PCR and not during library preparation.

Hilo1: /* HOW SINGLE MOLECULE SEQUENCING WORKS */

2025-04-23T18:24:58Z

HOW SINGLE MOLECULE SEQUENCING WORKS

← Older revision		Revision as of 18:24, 23 April 2025
Line 39:		Line 39:

	== [http://www.pacb.com/products/smrt-technology/ HOW SINGLE MOLECULE SEQUENCING WORKS ]==		== [http://www.pacb.com/products/smrt-technology/ HOW SINGLE MOLECULE SEQUENCING WORKS ]==
	The ~~Sequel~~ works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. A PacBio library with a single polymerase is loaded into each well. When a nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>		The Revio works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. A PacBio library with a single polymerase is loaded into each well. When a nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected. <BR><BR>
	The ~~Sequel~~ is able to monitor ~~>500,000~~ incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>		The Revio is able to monitor ~10 M incorporations occurring simultaneously on a SMRTcell. Each run creates a “movie” of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files. <BR><br>
	Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 20 hr movie. A mode called Continuous Long Read Sequencing simply sequences a subread sequenced over the length of the run, either 10 or 20 hours. In CLR mode, rare lengths of 175 kbp have been seen, but in the Center, it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.		Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 20 hr movie. A mode called Continuous Long Read Sequencing simply sequences a subread sequenced over the length of the run, either 10 or 20 hours. In CLR mode, rare lengths of 175 kbp have been seen, but in the Center, it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.