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Biomiga 96-well Tissue RNA Kit - R6811-00

  • Model: R6811-00
  • Manufactured by: Biomiga

£160.00

Description :

Introduction

 

The EZgeneTM 96 Well Tissue RNA Kit provides an easy and fast method for isolating total RNA from animal tissues, including some difficult fibrous tissues such as skeletal muscle, heart and aorta tissue. 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. This kit purifies up to 50 µg of total RNA from animal tissues. The purified RNA is ready for RT-PCR.

 

Storage and stability

 

DNase I (optional) and Proteinase K should be stored at -20°C. All other components can be stored at room temperature. All kit components are guaranteed for 1 year from the date of purchasing. 

Kit contents

 

Catalog#

R6811-00

R6811-01

R6811-02

Preps

1 x 96

4 x 96

20 x 96

96-well RNA Plate

1

4

20

96  Deep-well Plate

3

12

60

96-well Collection Plate (500 µL)

1

4

20

Buffer LY

40 mL

160 mL

2 x 400 mL

Buffer RB

125 mL

500 mL

5 x 500 mL

RNA Wash Buffer

40 mL

2 x 80 mL

10 x 80 mL

DEPC-Treated ddH2O

25 mL

100 mL

500 mL

Proteinase K

28 mg

4  x 28 mg

550  mg

DNase I  (RNase-free, 2U/µL)**

600 U

(300 µL)

2400 U

(1200 µL)

5 x 2400 U

(6 mL)

1 x DNase I Buffer**

8 mL

32 mL

5x32 mL

DNase Stop Buffer

24 mL

100 mL

5 x 100 mL

User Manual

1

1

1

*Add 160 mL (R6811-00) or 2 x 320 mL (R6811-01) or 10 x 320 mL (R6811-02) 96-100% ethanol to RNA Wash Buffer before use.

** DNase I and 10 x DNase I Buffer are shipped separately by ice.

 

Safety information

Buffer LY contains chaotropic salts, which may form reactive compounds when combines with bleach. Do not add bleach or acidic solutions directly to the preparation waste, wear gloves and protective eyewear when handling.

Before starting

 

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

Important

  •  Determine the volume of Buffer LY to be used and add 20 µL of β-mercaptoethanol (β-ME) per 1 mL Buffer LY before use. Buffer LY contains β-ME can be stored at room temperature for up to 1 month.
  •  Crystals may form in Buffer LY, dissolve the precipitates at 37oC before use.
  •  Add 160 mL (R6811-00) or 2 x 320 mL (R6811-01) or 10 x 320 mL (R6811-02) 96-100% ethanol to RNA Wash Buffer before use.
  •  Reconstitute Proteinase K with 1.1 mL (R6811-00) or 4 x 1.1 mL (R6811-01) or 22 mL (R6811-02) DEPC-Treated ddH2O, Vortex vial briefly and aliquot into adequate portions. Store aliquots at -20 oC.
  •  Add 36 mL (R6811-00) or 150 mL (R6811-01) or 5 x 150 mL (R6811-02) 100% ethanol to DNase Stop Buffer before use.
  •  All centrifuge steps must be carried out at 22 oC.

Materials supplied by users

  •  Centrifuge with swinging-bucket capable of 4000 g
  •  Rotor and adapter for 96-well plate
  •  Water bath or heat block preset at 55 oC
  •  100% ethanol
  •  b-mercaptoethanol
  •  RNase-free pipette tips

Troubleshooting Guides

 

Problem

Cause

Suggestion

Little or no RNA eluted

RNA remains on the membrane

  • l Repeat elution
  • l Pre-heat DEPC-water to 70℃prior to elution
  • l Incubate for 5 min with water prior to elution

Overloaded sample

Reduce quantity of starting material

Clogged wells

Incomplete lysis

  • l Mix thoroughly after addition of Buffer LY
  • l Reduce amount of starting material

Degraded RNA

Source

  • l Do not freeze and thaw sample more than once
  • l Follow protocol closely, and work quickly
  • l Low concentration of virus in the sample

RNase contamination

  • l Ensure not to introduce RNase during the procedure
  • l Check buffers for RNase contamination

Problem in

downstream

applications

Salt carry-over during elution

  • l Ensure RNA Wash Buffer has been diluted with 4 volumes of 100% ethanol as indicated on bottle
  • l 1 X RNA Wash Buffer must be stored at room temperature
  • l Repeat wash with RNA Wash Buffer

Inhibitors of PCR

  • l Use less starting material
  • l Prolong incubation with Buffer LY to completely lyse cells

DNA

contamination

 

Make sure to perform RNase-free DNase Digestion correctly

Abnormal OD

reading on

A260/A280

 

DEPC residue

remains in

DEPC-water

  • l Remove DEPC by Autoclave
  • l Use 10mM Tris-HCl, not the DEPC water to dilute the sample before measuring purity

Related EZgeneTM products

 

Catalog #

Product Name

Preps

Price $

R6311-01

Tissue RNA kit

50

150.00

R6311-02

Tissue RNA kit

250

650.00

R1011-01

RNASecure Solution

50 mL

45.00

R1011-02

RNASecure Solution

250 mL

150.00

R6312-01

Tissue RNA midi kit

10

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

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

R6812-01

96-well blood RNA kit

4x96

780.00

R6812-02

96-well blood RNA kit

20x96

3500.00


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