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Biomiga Blood RNA Maxi Kit - R6414-01

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Description :

Introduction

The EZgeneTM Blood total RNA kit provides an easy and fast method for isolating total RNA from white blood cells within 30 min. Only trace genomic DNA exists in the purified RNA, which can be eliminated by DNase I treatment (See detail in the protocol) when it is necessary.

Storage and Stability

DNase I (optional) should be stored at -20 ℃. All other components can be stored at room temperature. All kit components are guaranteed for 12 months from the date of purchasing.

Before Starting

Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each steps.

Important

  •  Add 1% volume of β-mercaptoethanol to Buffer LY before use and store at 4℃.
  •  Add 8 mL(R6411-00) or 80 mL (R6411-01) or 96 mL (R6411-02) 100% ethanol to RNA Wash Buffer before use.
  •  Optional: Add 800 µL (R6411-00) or 9.6 mL (R6411-01) or 48 mL (R6411-02) 100% ethanol to DNase Stop Buffer before use.
  •  Red Blood Cell Lysis Solution is supplied as 10 x concentrate. Calculate the amount of buffer to be used and mix one part of 10 x Red Blood Cell Lysis Solution with 9 part of ddH2O before use.

Materials supplied by users

  •  Tabletop microcentrifuge and 1.5 mL sterile tubes.
  •  Vacuum manifold if use vacuum protocol.
  •  100% ethanol
  •  Optional: DNase I, DNase Buffer

Note: Perform all steps including centrifugation at room temperature

Trouble Shooting Guide

Problem

Possible reason

Suggested Improvement

Low A260/A280 ratios

Protein contamination

Do a Phenol: Chloroform extraction. Loss of total RNA (up to 40%) should be expected.

Low A260/A280 ratios

Guanidine Thiocyanate contamination

Add 2.5 volumes of ethanol and 0.1M NaCl (final concentration) to precipitate RNA. Incubate for 30 min at -20 ℃. Centrifuge at 10,000 g for 15 min at 4 ℃ Resuspend the RNA pellet in DEPC-treated water.

Low Yield

RNA in sample degraded

Freeze samples immediately in liquid nitrogen and store at -70 ℃ after collect it.

Low Yield

The binding capacity of the membrane in the spin column was exceeded

Use of too much tissue sample exceeding the binding capacity of spin column will cause the decreasing of total RNA yield.

Low Yield

Ethanol not added to buffer

Add ethanol to the RNA Wash Buffer and DNase Stop Buffer before purification.

Genomic DNA contamination

Too much total RNA sample was used in RT-PCR.

Reduce total RNA amount used in RT-PCR to 50-100 ng.

Genomic DNA contamination

The sample may contain too much genomic DNA.

Reduce the amount of starting tissue in the preparation of the homogenate. Most tissues will not show a genomic DNA contamination problem at 30 mg or less per prep.

Reduce cell numbers to 1-2x106 or increase buffer volume and do multiple loadings to column.

Related EZgeneTM Products

Catalog #

Product Name

Preps

Price $

R6311-01

Tissue RNA kit

50

150.00

R6311-02

Tissue RNA kit

250

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R6312-01

Tissue RNA midi kit

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80.00

R6312-02

Tissue RNA midi kit

20

160.00

R6314-01

Tissue RNA maxi kit

10

120.00

R6314-02

Tissue RNA maxi kit

25

270.00

R6315-01

Tissue RNA mega kit

2

120.00

R6315-01

Tissue RNA mega kit

10

580.00

R6811-01

96-well tissue RNA kit

4x96

780.00

R6811-02

96-well tissue RNA kit

20x96

3300.00

R6411-01

Blood RNA mini kit

50

150.00

R6411-02

Blood RNA mini kit

250

680.00

R6412-01

Blood RNA midi kit

10

80.00

R6412-02

Blood RNA midi kit

25

180.00

R6414-01

Blood RNA maxi kit

10

120.00

R6414-02

Blood RNA maxi kit

25

270.00

R6812-01

96-well blood RNA kit

4x96

780.00

R6812-02

96-well blood RNA kit

20x96

3500.00


  • Model: R6414-01
  • Manufactured by: Biomiga

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