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96 Well Bacteria DNA Kit (96GBB04, 96GBB10)

DNA Extraction/Genomic DNA Purification

The Presto™ 96 Well gDNA Bacteria Kit is optimized for high-throughput genomic and viral DNA purification from Gram (-) negative and Gram (+) positive bacterial cells. Gram+ Buffer, when combined with Lysozyme, will efficiently lyse bacterial cell walls consisting of the peptidoglycan layer. Proteinase K and chaotropic salt are used to further lyse cells and degrade protein, allowing DNA to easily bind to the glass fiber matrix of the binding plate. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. Phenol/chloroform extraction or alcohol precipitation is not required and the purified genomic DNA is ready for use in a variety of downstream applications.

DOCUMENTS

COMPLETE PROTOCOL

DESCRIPTION

Specifications (Cat. # 96GBB04, 96GBB10)

High-throughput bacteria DNA extraction
High Yield: up to 5 μg of genomic DNA per well
High Purity gDNA: A260/A280 >1.9
DNA Size: 20-40 kb
Samples: Gram (+) Positive and Gram (-) Negative bacterial cells (up to 5 x 10⁸)
Convenient: Includes Gram+ Buffer for preparing Lysozyme solutions to speed up sample preparation when purifying DNA from Gram (+) positive bacteria
96 Well Binding Plates: glass fiber membrane optimized for genomic DNA extraction (sterilised to remove bacteria contamination)
Elution volume: 100-200 µl
Storage: dry at room temperature (15-25°C), Lysozyme is shipped at room temperature and should be stored at -20°C for extended periods

APPLICATIONS

PCR, Restriction Endonuclease Digestions, Array, Southern Blots and various other enzymatic reactions

COMPONENTS

Proteinase K
Gram+ Buffer
GT Buffer
GB Buffer
W1 Buffer
Wash Buffer
Lysozyme
Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
Presto™ gDNA 96 Well Binding Plates
Microtubes (Racked)
Microtubes (8-strip)
Caps for Microtubes (8-strip)
96 Deep Well Plates
Adhesive Film

QUALITY CONTROL

The quality of the Presto™ 96 Well gDNA Bacteria Kit is tested on a lot-to-lot basis by isolating DNA from Escherichia coli (5×10⁸) culture. 5 µl from a 100 µl eluate of purified DNA is analyzed by electrophoresis on a 1% agarose gel.

Presto™ 96 Well gDNA Bacteria Kit Functional Test Data (Gram (-) Negative Bacteria

Figure 1 .Genomic DNA was extracted from bacteria cells using the Presto™ 96 Well gDNA Bacteria Kit. The purified genomic DNA was eluted in 100 µl of Elution Buffer and 10 µl aliquots of the final sample (chosen from 6 random wells) were analyzed by electrophoresis on a 1% agarose gel.

M = Geneaid 1 Kb DNA Ladder
1-3: 5 x 10⁸ Escherichia coli cells, 4-6: 5 x 10⁸ Bacillus subtilis cells

Sample

ng/µl

260/280

Total Yield

Test 1

49.2

1.80

3.9 µg

Test 2

56.2

1.79

4.5 µg

Test 3

55.8

1.80

4.5 µg

Test 4

22.4

1.87

1.8 µg

Test 5

19.9

1.84

1.6 µg

Test 6

20.4

1.80

1.6 µg

Plasmid DNA Midi and Maxi Kits Specifications

Specifications	gSYNC™ DNA Extraction Kit	Genomic DNA Mini Kit Blood Cultured Cell	Genomic DNA Mini Kit Tissue	Genomic DNA Mini/Maxi Kit Plant	Presto™ Mini gDNA Bacteria Kit	Presto™ Mini gDNA Yeast Kit	Geneius™ Micro gDNA Extraction Kit	Presto™ Buccal Swab gDNA Extraction Kit
Cat. #	GS100 GS300	GB100 GB300	GT050 GT100 GT300	GP100 GPM010 GPM025	GBB100/101 GBB300/301	GBYB100 GBYB300	GMB100 GMB300	GSK100 GSK300
Format	Spin Column	Spin Column	Spin Column	Spin Column	Spin Column	Reagent System	Spin Column	Spin Column
Binding Capacity	30 µg	30 µg	30 µg	50 µg (Mini) 500 µg (Maxi)	50 µg	N/A	50 µg	50 µg
Sample	200 µl blood	300 µl blood	30 mg animal tissue	Mini: 100 mg plant tissue	1x10⁹ bacterial cells	2x10⁸ yeast cells	1-100 µl blood	buccal swabs
	25 mg animal tissue	200 µl buffy coat		Maxi: 1 g plant tissue	200 µl blood		FTA® Cards
	1x10⁷ cultured cells	1x10⁷ cultured cells			1 ml urine, plasma		1 ml urine
	hair	2x10⁸ yeast/fungus					1-100 µl saliva
Micropestle	N/A	N/A	Yes	N/A	N/A	N/A	N/A	N/A
Filter Column	N/A	N/A	N/A	Yes	N/A	N/A	N/A	Yes
Enzyme	Proteinase K	N/A	Proteinase K	N/A	Lysozyme Proteinase K	Lyticase or Zymolase	Proteinase K	Proteinase K
Typical Yield	5 µg from 200 µl blood	4-10 µg from 300 µl blood	10-40 µg from 20 mg tissue	3-5 µg from 100 mg Arabidopsis leaf (Mini)	37 µg from 1x10⁹ E. coli	10-15 µg from 2x10⁸ S. cerevisiae cells	300 ng from 6 mm blood spots	2 µg per swab
	10-40 µg from 20 mg tissue			80 µg from 500 mg Lemon leaf (Maxi)	10-15 µg from 1x10⁹ Bacillus subtilis
Elution Volume	30-100 µl	30-200 µl	30-200 µl	30-200 µl (Mini) 1 ml (Maxi)	30-200 µl	50-100 µl	30-100 µl	50-100 µl
Operation Time	<20 minutes	<25 minutes	30 minutes	<30 minutes	<30 minutes	<40 minutes	<20 minutes	<20 minutes

RELATED PRODUCTS

Product Name	RXNS	Catalogue Number
gSYNC™ DNA Extraction Kit (Tissue/Blood)	100, 300	GS100/300
Genomic DNA Mini Kit (Blood/Cultured Cell)	100, 300	GB100/300
Genomic DNA Maxi Kit (Plant)	10, 25	GPM010/25
Genomic DNA Maxi Kit (Blood/Cultured Cell)	10, 25	GDM10/25
Genomic DNA Mini Kit (Tissue)	50, 100, 300	GT050/100/300
Genomic DNA Mini Kit (Plant)	100	GP100
Geneius™ Micro gDNA Extraction Kit	100	GMB100/300
Presto™ Buccal Swab gDNA Extraction Kit	100	GSK100/300
Presto™ Mini gDNA Yeast Kit	100, 300	GBYB100/300
Presto™ Mini gDNA Bacteria Kit	100, 300	GBB100/101/300/301
Presto™ 96 Well Blood Genomic DNA Extraction Kit	4 x 96, 10 x 96	96GBP04/10
Presto™ 96 Well Plant Genomic DNA Extraction Kit	4 x 96, 10 x 96	96GPP04/10
Presto™ 96 Well gDNA Bacteria Kit	4 x 96, 10 x 96	96GBB04/10

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