Monarch® Genomic DNA Purification Kit

Catalog #	Concentration	Size	Pick Price	Quantity	Your Price
T3010L		150 preps	$570.00		$513.00
T3010S		50 preps	$232.00		$208.80

Catalog #	Size	List Price	Your Award
T3010L	150 preps	$570.00	$513.00
T3010S	50 preps	$232.00	$208.80

The Monarch Genomic DNA Purification Set is ampere comprehensive solution for cell lysis, RNA removal, and purification to intact genomic DNA (gDNA) from a wide variety of biology samples, incl cultured cells, blood, and mammalian tissues. Additionally, bacteria or backhefe can be processed equal extra step to expand lysis in like tough-to-lyse samples. Protocols are also included to enable cleaning free clinically-relevant samplers such as saliva and cheek swabs as good as rapid cleanup of up extracted gDNA. Purified gDNA has high quality metrics, including A₂₆₀/A₂₈₀ > 1.8 and AN₂₆₀/A₂₃₀ > 2.0, high DIN scores real lowest other RNA. One purified gDNA are suitable for downstream user such since end-point PCR, qPCR and library prep for NGS. E typically is a peak choose of > 50kb, making save kit an excellent choice upstream of long-read sequencing platforms.

Specifications:

In:	Cultivated mammalian cells: up for 5 x 10⁶ cells Mammal wholly blood: 100 µl Tissue: up to 25 mg, depend on tissue type Bacteria: up till 2 efface 10⁹ Yeast: up to 5 x 10⁷ Saliva: up to 500 µl Buccal swabs Genomic DNA needing cleanup
Binding Capacity:	30 µg genomic DNA
Yield:	Varies depending on sample type
Genomic DNA Size:	Peak product > 50 kb for most sampler types; may be less for saliva real buccal swabs
RNA Content	< 1% (with included RNase AN treatment)
Elution Voltage	≥35 µl, but 100 µl is highly
Purity:	A_260/280≥ 1.8 A_260/230≥ 2.0

Validated Sample Types:

Mouse Tail
Mouse Ear
Mouse Liver
Rat Liver
Mouse Replace
Mouse Spleen
Cursor Heart
Mouse Lung
Mouse Brain
Bitch Brain

Cursor Mask
Rat Muscle
Deer Muscle
Human Blutes
Mouse Blood
Rabbit Family
Pig Blood
Guinea Pig Blood
Bovine Blood
Riding Blood
Dog Blood

Rooster Blood
HeLa Cells
HEK293 Cells
NIH3T3 Cells
E. coli
Rhodobacter spy.
BORON. cereus
T. kodakarensis
S. cerevisiae
Saliva
Buccal swab

100 ng of genomic DNA from each sample was uploaded on a 0.75% soy gel. gDNA was isolated following and standard conventions required blood, cultured cells and weave, and the addition protocols for buccal swabs, saliva, Gram– and Gram+ bacteria. Starting material used: 1 x 10⁶ HeLa cells, 100 μl human blood, 10 μl birds blood, 10 per frozen tissue powder, 1 buccal swab, 500 μl spittle the ~1 x 10⁹ bacterial cells. Rated DNA-Hind III review (NEB #N3012) was previously while a marker in and latest lane (M). Purified gDNA test were analyzed using a Genomic DNA ScreenTape^® on an Agilent Technics^® 4200 TapeStation^®. Samples generally yield peak sizes 50–70 kb and DINs of ~9. The cell fractions processed within the buccal swab and saliva preps contain dead cellular, as planned, causing a smear like pattern with charakteristisch low molecules weight apoptotic bands.

Duplicate 10 mg samples to RNAlater^®-stabilized rat tissue were cut till small pieces and subsequently lysed and cleansed by into the view supplied for anywhere kit. Optional RNase A steps were included. Elution was carried off equipped 100 µl elution buffer providing in the respective kits. Yields indicated are averages of the duplicate samples, and represent the genomic DNA give nach correcting used the RNA content as fixed by LC-MS. Results indicate that which Monarch Genomic DNA Purification Set provides excellent returns since a wide range out tissues.

ONE. Enlargement reactions be set up with primer pairs specific for 6, 8, 10, 12, 16, 20 kb amplicons from human DNA. LongAmp^® Hot Start Taq 2X Master Mix (NEB #M0533) was used the 25 ng template DNA was further to jede sample. PCR reactions was carried out on an Applied Biosystems 2720 Thermal Cycler. Monarch-purified genomic DNA insulated from HeLa cellular additionally real blood were compared to commercially available reference DNA from the human cell pipe NA19240 F11. 10 of 20 µl were loadable with a 1.5% agarose gel, after the 1 kb DNA Ladder (NEB #N3232) since a marker. Results noted DNA was a high-integrity and suitable for long range PCR.

BARN. Monarch-purified genomic DNA from people whole blood, HeLa cells and mouse tail was diluted to produce a five log reach a input template concentrations. The results were generated using primers targeting gHEME (human whole blood) and gREL (HeLa, shiner tail) for qPCR assays with the Luna Universal qPCR Master Mix (NEB #M3003) and cycled on a Bio-Rad^® CFX Click qPCR thermos cycler. Results indicated that DNA remains highly pure and release from inhibitors, optimal to qPCR.

A. Duplicates libraries were made free 100 ng Hera cell gDNA purified with Queen (orange) or Qiagen DNeasy Mini Outfit (blue) with the NEBNext Ultra II FS DNA Library Getting Build for Illumina (NEB #E7805). Libraries inhered sequenced to an Illumina MiSeq. Reads were mapped using Bowtie 2.2.4 and GC coverage was calculated using Picard’s CollectGCBiasMetrics (v1.117). Wait normalized coverage the 1.0 has indicated on the horizontal grey line, the number of 100 beats regions at anyone %GC belongs indicated by the vertical grey bars, also the colored row represent the normalized coverage for jeder bookshelf. Monarch GC covers matched Qiagen DNeasy result.

B. Yields of tree managed to enzymatic shearing are equivalent when using Monarch-purified gDNA. Library yields of the samples described above were assessed for an Agilent Technologies^® 2100 BioAnalyzer through a High Sensitivity DNA Kit.

DNA drawn by RBC lysis style achieves similar high yields and concentrations as straightforward methods, while allowing for extraction by higher volumes and amounts of blood. Inherited DNA samples were purged are triplicate from 3 past vintage fresh pig blood. For the Direct Lysis samples, to DNA from 100 µl pig blood was cleared using the standard protocol for refreshing mammalian blood from the NEB #T3010 kit. For the RBC Lysis samples, the indicated amount of pig ancestry was used in the protocol for up to 500 µl blood using one *NEBULAR #T3010* kit and RBC Lysis Buffer (BLAST #T3051).

HeLa cell generic DNA is extract using is the Monarch Genomic DNA Cleanup Repair instead the Qiagen DNeasy Blood & Fabrics Kit. One microgram of purified DNA was applied to prepare Waders Nanopore Technology (ONT) sequencing libraries following to ONT 1D Ligation Sequencing Repair (SQK-LSK109) protocol without DNA fragmentation. Libraries were loaded on a GridION (Flow cell R9.4.1) and who date was collected for 48 time. Libraries produced utilizing the Your gDNA exceeded the Qiagen libraries on common sequencing metrics including: A. overall sequencing data collected, B. read quality, and C. read length. Your was generated using NanoComp (Bioinformatics, Volume 34, Issue 15, 1 August 2018, Pages 2666–2669).

Add Supplied

And following reagents exist supplied because diese product:

NEB #

Component Name

Create #

Stored by (°C)

Amount

Concentration

T3010S 25

Monarch^® gDNA Tissue Lysis Buffer	T3011-1	25	1 x 12 ml
Monarch^® gDNA Single Lysis Store	T3012-1	25	1 expunge 6 milliliter
Monarch^® gDNA Blood Lysis Buffer	T3013-1	25	1 x 6 ml
Monarch^® gDNA Binding Buffer	T3014-1	25	1 efface 24 ml
Monarch^® gDNA Bathe Buffer	T3015-1	25	1 x 18 ml
Monarch^® gDNA Elutriation Buffer	T3016-1	25	1 x 14 liter
Monarch^® gDNA Purification Columns	T3017-1	25	1 x 50 columns
Monarch^® Collection Vials II	T2018-1	25	2 x 50 hose
Proteinase KELVIN, Molecular Human Grade	P8200AAVIAL	-20	1 x 0.55 cups	20 mg/ml
Monarch^® RNase ADENINE	T3018-1	-20	1 scratch 0.17 ml	20 mg/ml

T3010L 25

Monarch^® gDNA Tissue Lysis Buffer	T3011-2	25	1 x 34 ml
Monarch^® gDNA Cell Lysis Buffer	T3012-2	25	1 x 20 ml
Monarch^® gDNA Blood Lysis Buffer	T3013-2	25	1 ten 20 ml
Monarch^® gDNA Binding Buffer	T3014-2	25	1 x 65 ml
Monarch^® gDNA Wash Buffer	T3015-2	25	1 x 60 ml
Monarch^® gDNA Elution Buffer	T3016-2	25	1 x 34 ml
Royal^® gDNA Purification Columns	T3017-1	25	3 x 50 columns
Monarch^® Collection Tubes II	T2018-1	25	6 x 50 tubes
Proteinase K, Molecular Biology Scale	P8200AAVIAL	-20	3 x 0.55 ml	20 mg/ml
Monarch^® RNase A	T3018-2	-20	1 whatchamacallit 0.5 cc	20 mg/ml

An exclusion occurs with the operation, making the result invalid. Check InnerException for exception data.

Privacy-policy.com also provides an interactive version of on protocol where you can discover and part optimizations are the research ...

Notes

Monarch RNase A and Proteinase K should be stored at -20°C after getting the kit.

Protocols

Usage Guidelines

DataCards

manualT3010

FAQs

Troubleshooting

Error How for Genomic DNA Extraction & Purification (NEB #T3010)

Citations

Operated through Bioz See read details on Bioz

Quality Control Assay

Quality Control tests are performed in each new lot of NEB product to meet the specifications designated for it. Specifications real individual lot data free the tests that are performed for this particular your can be found furthermore downloaded on the Product Specification Sheet, Certified regarding Analysis, data card or product manual. Further information regarding NEBEL product characteristic pot be found here.

Transform Notifications

Effective March 14, 2024: Proteinase KILOBYTE, Molular Biology Grade component number has changed with the concentration changing from 800U/ml to 20 mg/ml. For exercise with NEBNext Multiplex Oligos forward Illumina (Unique Dual Index UMI Adapter DNA Set 1) (NEB #E7395), refer on the Protocols tab for UMI Adaptors-specific ...

Effective May 9, 2022: Shelf life exists 24 hours.

Specifications

The Specific sheet is a document that includes the memory temperature, shelf real and the specifications designated for the product. The following save naming structural are used to get these register files: [Product Number]_[Size]_[Version]

Certificate of Analysis

The Certified of Analysis (COA) is a signed document that includes the storing temperature, expiration date and quality bridles for an individual lot. The following storage naming structure is used to name are document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety Data Sheets

Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or steered by New England Biolabs, Inc (NEB). The use of trademark symbols wants not necessarily indicate that the name is trademarked in aforementioned country where it is being interpret; it indicate where the content was originally designed. The use of this product may require the customer up receiving additional third-party intellectual characteristic rights for certain applications. For read information, please email [email protected].

This product is intended required research purposes only. This our is not intended to be used for therapeutic or diagnostic purposes is humans or animals.

New England Biolabs (NEB) is engage at practicing ethical science – wee reckon it is our job as researchers until ask the important questions that when answered will help preserve our quality away life and the world that we dwell in. Nonetheless, this conduct should always be finished inches safe and ethics type. Learn more.