BioMicroCenter:DNA LIB - Revision history

172.18.0.1: /* Nextera XT */

2025-05-02T14:53:40Z

Nextera XT

← Older revision		Revision as of 14:53, 2 May 2025
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	Nextera uses a modified Tn5 transposase to simultaneously fragment intact genomic DNA and tag it with Illumina adapters. A limited number of PCR steps are required to generate complete ~~Illumina~~ libraries. This very simple method makes this preparation very popular for automation and in vivo methods such as ATAC-Seq.<br><br>		Nextera uses a modified Tn5 transposase to simultaneously fragment intact genomic DNA and tag it with Illumina adapters. A limited number of PCR steps are required to generate complete NGS short read libraries. This very simple method makes this preparation very popular for automation and in vivo methods such as ATAC-Seq.<br><br>

	[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna.html Original Nextera] requires 50 ng of input material and produces the most complex libraries. Products can be sized by [[BioMicroCenter:PippinPrep\|Pippin]] and are suitable for all applications. [https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-xt-dna.html NexteraXT] uses less input material (1 ng) and less enzyme, making it less expensive, but only crude size fractionation using single-pass SPRI selection can be done. An automated version of Nextera XT is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-Throughput DNA preparation page]].		[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna.html Original Nextera] requires 50 ng of input material and produces the most complex libraries. Products can be sized by [[BioMicroCenter:PippinPrep\|Pippin]] and are suitable for all applications. [https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-xt-dna.html NexteraXT] uses less input material (1 ng) and less enzyme, making it less expensive, but only crude size fractionation using single-pass SPRI selection can be done. An automated version of Nextera XT is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-Throughput DNA preparation page]].

172.18.0.1: /* NEB Ultra II */

2025-05-02T14:50:49Z

NEB Ultra II

← Older revision		Revision as of 14:50, 2 May 2025
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	The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and ~~the i5 and i7~~ anchors. Completed libraries can be size selected using SPRI beads or BluePippin.		The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and NGS short read anchors. Completed libraries can be size selected using SPRI beads or BluePippin.
	\|		\|
	[[image:NEB_Data.jpg\|thumb\|right\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]		[[image:NEB_Data.jpg\|thumb\|right\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]

Hilo1: /* Nextera Flex */

2025-04-30T13:06:19Z

Nextera Flex

← Older revision		Revision as of 13:06, 30 April 2025
Line 68:		Line 68:
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	~~[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna-flex.html~~ Nextera ~~FLEX]~~ uses bead-linked transposome chemistry for on-bead tagmentation. This provides steric inhibition which increases the size of the inserts in the library. At and above 100 ng DNA input, the beads are saturated and libraries are self-normalized. An automated version of Flex is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-Throughput DNA preparation page]].		Nextera Flex, also known as Illumina DNA Prep, uses bead-linked transposome chemistry for on-bead tagmentation. This provides steric inhibition which increases the size of the inserts in the library. At and above 100 ng DNA input, the beads are saturated and libraries are self-normalized. An automated version of Flex is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-Throughput DNA preparation page]].

Hilo1: /* NEB Ultra II */

2025-04-30T13:05:30Z

NEB Ultra II

← Older revision		Revision as of 13:05, 30 April 2025
Line 44:		Line 44:
	The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.		The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.
	\|		\|
	[[image:NEB_Data.jpg\|thumb\|right~~\|500px~~\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]		[[image:NEB_Data.jpg\|thumb\|right\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]

	\|}		\|}

Hilo1: /* NEB Ultra II */

2025-04-30T13:04:53Z

NEB Ultra II

← Older revision		Revision as of 13:04, 30 April 2025
Line 44:		Line 44:
	The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.		The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.
	\|		\|
	[[image:NEB_Data.jpg\|right\|~~350px~~\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]		[[image:NEB_Data.jpg\|thumb\|right\|500px\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]

	\|}		\|}

Hilo1: /* NEB Ultra II */

2025-04-30T13:04:36Z

NEB Ultra II

← Older revision		Revision as of 13:04, 30 April 2025
Line 44:		Line 44:
	The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.		The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.
	\|		\|
	[[image:NEB_Data.jpg~~\|thumb~~\|right\|350px\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]		[[image:NEB_Data.jpg\|right\|350px\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]

	\|}		\|}

Hilo1: /* Nextera XT */

2025-04-25T18:13:29Z

Nextera XT

← Older revision		Revision as of 18:13, 25 April 2025
Line 97:		Line 97:
	Nextera uses a modified Tn5 transposase to simultaneously fragment intact genomic DNA and tag it with Illumina adapters. A limited number of PCR steps are required to generate complete Illumina libraries. This very simple method makes this preparation very popular for automation and in vivo methods such as ATAC-Seq.<br><br>		Nextera uses a modified Tn5 transposase to simultaneously fragment intact genomic DNA and tag it with Illumina adapters. A limited number of PCR steps are required to generate complete Illumina libraries. This very simple method makes this preparation very popular for automation and in vivo methods such as ATAC-Seq.<br><br>

	[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna.html Original Nextera] requires 50 ng of input material and produces the most complex libraries. Products can be sized by [[BioMicroCenter:PippinPrep\|Pippin]] and are suitable for all applications. [https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-xt-dna.html NexteraXT] uses less input material (1 ng) and less enzyme, making it less expensive, but only crude size fractionation using single-pass SPRI selection can be done. An automated version of Nextera XT is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-~~Throuhgput~~ DNA preparation page]].		[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna.html Original Nextera] requires 50 ng of input material and produces the most complex libraries. Products can be sized by [[BioMicroCenter:PippinPrep\|Pippin]] and are suitable for all applications. [https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-xt-dna.html NexteraXT] uses less input material (1 ng) and less enzyme, making it less expensive, but only crude size fractionation using single-pass SPRI selection can be done. An automated version of Nextera XT is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-Throughput DNA preparation page]].

	Nextera works well with amplicons as well as gDNA. However, because two hits are required per molecule to create productive libraries, the ratio of reagent to fragment must be altered significantly to produce good libraries and large inserts may not be possible.		Nextera works well with amplicons as well as gDNA. However, because two hits are required per molecule to create productive libraries, the ratio of reagent to fragment must be altered significantly to produce good libraries and large inserts may not be possible.

Hilo1: /* Nextera Flex */

2025-04-25T18:13:15Z

Nextera Flex

← Older revision		Revision as of 18:13, 25 April 2025
Line 68:		Line 68:
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	[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna-flex.html Nextera FLEX] uses bead-linked transposome chemistry for on-bead tagmentation. This provides steric inhibition which increases the size of the inserts in the library. At and above 100 ng DNA input, the beads are saturated and libraries are self-normalized. An automated version of Flex is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-~~Throuhgput~~ DNA preparation page]].		[https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna-flex.html Nextera FLEX] uses bead-linked transposome chemistry for on-bead tagmentation. This provides steric inhibition which increases the size of the inserts in the library. At and above 100 ng DNA input, the beads are saturated and libraries are self-normalized. An automated version of Flex is available. For more information, please look at the [[BioMicroCenter:DNA_HTL\|High-Throughput DNA preparation page]].

Hilo1: /* Nextera XT */

2025-04-25T18:12:45Z

Nextera XT

← Older revision		Revision as of 18:12, 25 April 2025
Line 101:		Line 101:
	Nextera works well with amplicons as well as gDNA. However, because two hits are required per molecule to create productive libraries, the ratio of reagent to fragment must be altered significantly to produce good libraries and large inserts may not be possible.		Nextera works well with amplicons as well as gDNA. However, because two hits are required per molecule to create productive libraries, the ratio of reagent to fragment must be altered significantly to produce good libraries and large inserts may not be possible.
	\|		\|
	[[Image:Nextera Illumina fig1.~~gif~~\|thumb\|right\|260px\|Standard Nextera workflow.]]		[[Image:Nextera Illumina fig1.jpg\|thumb\|right\|260px\|Standard Nextera workflow.]]
	\|}		\|}

172.18.0.1: /* NEB Ultra II */

2025-04-25T18:06:53Z

NEB Ultra II

← Older revision		Revision as of 18:06, 25 April 2025
Line 44:		Line 44:
	The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.		The BioMicro Center utilizes the [https://www.neb.com/products/e7645-nebnext-ultra-ii-dna-library-prep-kit-for-illumina#Product%20Information NEB UltraII] kit for standard library preparation. This kit is extremely robust and can make libraries from <1ng of input material. It is also amenable to miniaturization and is available as a high-throughput prep as well. NEB Ultra II is based on standard LM-PCR based chemistry. DNA fragments are repaired, A-tailed and ligated to adapters. The ligated products are then amplified by PCR to add indexes and the i5 and i7 anchors. Completed libraries can be size selected using SPRI beads or BluePippin.
	\|		\|
	[[image:NEB_Data.~~png~~\|thumb\|right\|350px\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]		[[image:NEB_Data.jpg\|thumb\|right\|350px\|NEBNext Ultra II produces the highest rates of conversion to adaptor-ligated molecules from a broad range of input amounts.]]

	\|}		\|}