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Blood/Cell RNA Mini Kit (RBD050, RBD100, RBD300) with DNase

RNA Extraction/Total RNA

The RNA Kit (Blood/Cultured Cell) is a RNA Isolation Kit designed for total RNA purification from fresh whole human blood and cultured cells. This RNA Kit will purify RNA from blood within 20 minutes using an efficient RNA miniprep system. Detergents and chaotropic salt are used to lyse cells and inactivate RNase. RNA in the chaotropic salt is bound by the glass fiber matrix of the RNA spin column and once any contaminants have been removed, using the Wash Buffer (containing ethanol), the purified total RNA is eluted by RNase-free Water and ready for use in various downstream applications. Each Geneaid RNA Kit includes individually packaged certified DNase and RNase-free spin columns and collection tubes and DNase I for DNA sensitive downstream applications.

DOCUMENTS

COMPLETE PROTOCOL

DESCRIPTION

Specifications (Cat. # RBD050, RBD100, RBD300)

Purify total RNA within 20 minutes!
High Yield: up to 2 µg from whole blood, up to 30 µg from 1x10⁶ 293T cells
Sample Volume: up to 300 µl of whole blood, cultured cells (up to 5 x 10⁶)
Spin Columns: glass fiber membrane optimized for total RNA extraction
Individually packaged RNA spin columns and collection tubes, certified RNase and DNase-free
Storage: dry at room temperature (15-25°C), DNase I is shipped at room temperature and should be stored at -20°C

APPLICATIONS

RT-PCR, Northern Blotting, Primer Extension, mRNA Selection, cDNA Synthesis

COMPONENTS

RBC Lysis Buffer
RB Buffer
DNase I
DNase I Reaction Buffer
W1 Buffer
Wash Buffer
RNase-free Water
RB Columns
2 ml Collection Tubes

QUALITY CONTROL

The quality of the Total RNA Mini Kit (Blood/Cultured Cell) is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. Total RNA from 300 μl of fresh whole human blood is extracted using the Total RNA Mini Kit (Blood/Cultured Cell). The purified RNA (2 μg with an A260/A280 ratio of 1.8-2.0) is quantified with a spectrophotometer and checked by electrophoresis.

Total RNA Mini Kit (Blood/Cultured Cell) Functional Test Data

Figure 1 .Total RNA from 1×10⁶ 293T cells was extracted using the Total RNA Mini Kit (Blood/Cultured Cell) and the equivalent competitors kit. The purified RNA was analyzed by electrophoresis on a 1% agarose gel.
Lane 1, 2 = Geneaid, Lane 3, 4 = Competitor, M = Geneaid 1 Kb DNA Ladder

Geneaid

Competitor

Yield (μg)

260/280

260/230

Yield (μg)

260/280

260/230

28.70

1.96

2.32

20.50

1.85

2.31

29.90

1.99

2.02

28.90

1.84

2.32

Figure 2 .Total RNA from 300 µl of whole human blood was extracted using the Total RNA Mini Kit (Blood/Cultured Cell) . The purified RNA was analyzed by electrophoresis on a 1% agarose gel.

M = Geneaid 1 Kb DNA Ladder

Yield (μg)

260/280

260/230

2.02

1.91

2.15

2.13

1.90

2.35

2.10

1.91

2.25

Figure 3 .Total RNA was purified from 10¹-10³ HeLa cells using the Total RNA Mini Kit Blood/Cultured Cell (2 replications of each cell number). The purified RNA was eluted with 25 μl RNase-free Water. cDNA synthesis was carried out with a 10 μl aliquot of purified RNA (2/5) using a Transcriptor First Strand cDNA Synthesis Kit (Roche) in a final volume of 20 μl. A Real-time PCR assay was then performed with 8 µl of synthesized cDNA (2/5) as template, primers (designed to amplify human ACTB gene), and Fast SYBR Green PCR Master Mix using the StepOnePlusTM Real-Time PCR system (Applied Biosystems). The results confirmed that RNA can be successfully extracted and detected from as low as 10¹ HeLa cells.

Legend
10³ HeLa Cells, 10² HeLa Cells, 10¹ HeLa Cells

Total RNA Extraction Kits Specifications

Specifications	Total RNA Mini Kit Blood Cultured Cell	Total RNA Mini Kit Tissue	Total RNA Mini Kit Plant	Presto™ Mini RNA Bacteria Kit	Presto™ Mini RNA Yeast Kit	miRNA Isolation Kit	GENEzol™ TriRNA Pure Kit
Cat. #	RBD050 RBD100 RBD300	RTD050 RTD100 RTD300	RPD050 RPD100 RPD300	RBBD050 RBBD100 RBBD300	RBYD050 RBYD100 RBYD300	RMI050	GZXD050 GZXD100 GZXD200
Format	Spin Column	Spin Column	Spin Column	Spin Column	Spin Column	Spin Column	Tri Reagent Spin Column
Binding Capacity	50 µg	50 µg	50 µg	50 µg	50 µg	50 µg	50 µg
Sample	blood, cells	animal tissue	plant tissue	bacterial cells	yeast cells	animal tissue, cells	blood,cells, tissue, body fluids, bacteria
Micropestle	N/A	Yes	N/A	N/A	N/A	Yes	N/A
Filter Column	N/A	Yes	Yes	N/A	N/A	N/A	N/A
Enzyme	Dnase I	Dnase I	Dnase I	Lysozyme Dnase I	Dnase I	N/A	Dnase I
Typical Yield	2-3 µg from 300 µl blood, 25-30 µg from 10⁶ cells	5-30 µg	5-30 µg	40-50 µg from 1x10⁹ bacterial cells	15-20 µg from 5x10⁷ yeast cells	5-30 µg	5-30 µg
Elution Volume	50 µl	50 µl	50 µl	50 µl	50 µl	50 µl	25-50 µl
Time	<20 min.	<15 min.	<20 min.	<20 min.	<20 min.	<30 min.	<15 min.

RELATED PRODUCTS

Product Name	RXNS	Catalogue Number
Total RNA Mini Kit (Blood/Cultured Cell)	50/100/300	RB050/100/300, RBD050/100/300
Total RNA Mini Kit (Tissue)	50/100/300	RT050/100/300, RTD050/100/300
Total RNA Mini Kit (Plant)	50/100/300	RP050/100/300, RPD050/100/300
Presto™ Mini RNA Bacteria Kit	50/100/300	RBB050/100/300, RBBD050/100/300
Presto™ Mini RNA Yeast Kit	50/100/300	RBY050/100/300, RBYD050/100/300
miRNA Isolation Kit	50	RMI050
GENEzol™ TriRNA Pure Kit	50/100/200	GZX050/100/200, GZXD050/100/200
GENEzol™ 96 Well TriRNA Pure Kit	4 x 96/10 x 96	96GZX04/10

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