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Qiagen RNEASY KITS User Manual
Page 99
User Manual - Page 99
For RNEASY KITS.
Page 99/100
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Contents
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RNeasy Min
i Handbook
10/2019
99
No
tes
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96
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100
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File type: PDF
File name: 25999280_rneasy-kits.pdf
File size: 806.78 KB
File Language: English
Pages: 100
Author: Qiagen
File created: 2019-11-06
Published: 2021-10-08
Updated: 2023-05-21
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Table of Contents
×
Kit Contents
4
Kit Contents
4
Storage
6
Storage
6
Intended Use
6
Intended Use
6
Safety Information
7
Safety Information
7
Quality Control
7
Quality Control
7
Introduction
8
Introduction
8
Principle and procedure
9
Principle and procedure
9
RNA purification using RNeasy technology
9
RNA purification using RNeasy technology
9
RNA stabilization using RNAprotect technology
10
RNA stabilization using RNAprotect technology
11
RNA stabilization using RNAprotect technology
11
Description of protocols
11
Purification of Total RNA from Animal Cells Using Spin Technology
11
Description of protocols
12
Description of protocols
12
Purification of Total RNA from Animal Cells Using Spin Technology
12
Purification of Total RNA from Animal Cells Using Vacuum/Spin Technology
12
Purification of Total RNA from Animal Cells Using Vacuum/Spin Technology
12
Stabilization of RNA in Harvested Animal Tissues
12
Stabilization of RNA in Harvested Animal Tissues
12
Purification of Total RNA from Animal Tissues
12
Purification of Total RNA from Animal Tissues
13
Purification of Total RNA from Animal Tissues
13
Purification of Total RNA from Yeast
13
Purification of Total RNA from Yeast
13
Purification of Total RNA from Plant Cells and Tissues and Filamentous Fungi
14
Purification of Total RNA from Plant Cells and Tissues and Filamentous Fungi
14
RNA Cleanup
14
RNA Cleanup
14
Automated purification of RNA on QIAcube Instruments
14
Automated purification of RNA on QIAcube Instruments
15
Automated purification of RNA on QIAcube Instruments
15
Equipment and Reagents to Be Supplied by User
16
Equipment and Reagents to Be Supplied by User
16
For all protocols
16
For all protocols
16
For RNA purification from animal cells
16
For RNA purification from animal cells
16
For RNA purification from animal cells using vacuum technology
16
For RNA purification from animal cells using vacuum technology
16
For RNA purification from animal tissues
17
For RNA purification from animal tissues
17
For RNA purification from yeast using enzymatic lysis
17
For RNA purification from yeast using enzymatic lysis
17
For RNA purification from yeast using mechanical disruption
17
For RNA purification from yeast using mechanical disruption
18
For RNA purification from plants and fungi
18
For RNA purification from plants and fungi
18
Suppliers of equipment for disruption and homogenization2F
18
Suppliers of equipment for disruption and homogenization2F
18
Suppliers of equipment for disruption and homogenization2F
18
Suppliers of equipment for disruption and homogenization2F
18
Suppliers of equipment for disruption and homogenization2F
18
Suppliers of equipment for disruption and homogenization2F
18
Suppliers of equipment for disruption and homogenization2F
18
Important Notes
19
Important Notes
19
Determining the amount of starting material
19
Determining the amount of starting material
19
Handling and storing starting material
20
Handling and storing starting material
20
Disrupting and homogenizing starting material
22
Disrupting and homogenizing starting material
22
Disruption and homogenization using the TissueLyser system
23
Disruption and homogenization using the TissueLyser system
23
Disruption and homogenization using rotorâstator homogenizers
24
Disruption and homogenization using rotorâstator homogenizers
24
Disruption using a mortar and pestle
24
Disruption using a mortar and pestle
25
Disruption using a mortar and pestle
25
Homogenization using QIAshredder homogenizers
25
Homogenization using QIAshredder homogenizers
25
Homogenization using a syringe and needle
25
Homogenization using a syringe and needle
25
Eliminating genomic DNA contamination
25
Eliminating genomic DNA contamination
26
Eliminating genomic DNA contamination
26
Protocol: Purification of Total RNA from Animal Cells Using Spin Technology
27
Protocol: Purification of Total RNA from Animal Cells Using Spin Technology
27
Determining the correct amount of starting material
27
Determining the correct amount of starting material
27
Important points before starting
28
Important points before starting
28
Things to do before starting
29
Things to do before starting
29
Procedure
29
Procedure
29
Protocol: Purification of Total RNA from Animal Cells Using Vacuum/Spin Technology
35
Protocol: Purification of Total RNA from Animal Cells Using Vacuum/Spin Technology
35
Determining the correct amount of starting material
35
Determining the correct amount of starting material
35
Important points before starting
35
Important points before starting
35
Things to do before starting
36
Things to do before starting
36
Procedure
36
Procedure
37
Procedure
37
Protocol: Stabilization of RNA in Harvested Animal Tissues
41
Protocol: Stabilization of RNA in Harvested Animal Tissues
41
Important notes about RNAprotect Tissue Reagent
41
Important notes about RNAprotect Tissue Reagent
41
Important points before starting
42
Important points before starting
42
Procedure
42
Procedure
43
Procedure
43
Protocol: Purification of Total RNA from Animal Tissues
45
Protocol: Purification of Total RNA from Animal Tissues
45
Determining the correct amount of starting material
45
Determining the correct amount of starting material
45
Important points before starting
46
Important points before starting
46
Things to do before starting
47
Things to do before starting
47
Procedure
47
Procedure
47
Protocol: Purification of Total RNA from Yeast
53
Protocol: Purification of Total RNA from Yeast
53
Disrupting yeast cells
53
Disrupting yeast cells
53
Determining the correct amount of starting material
53
Determining the correct amount of starting material
53
Important points before starting
55
Important points before starting
55
Things to do before starting
55
Things to do before starting
55
Procedure
56
Procedure
56
Protocol: Purification of Total RNA from Plant Cells and Tissues and Filamentous Fungi
62
Protocol: Purification of Total RNA from Plant Cells and Tissues and Filamentous Fungi
62
Determining the correct amount of starting material
62
Determining the correct amount of starting material
62
Important points before starting
62
Important points before starting
62
Things to do before starting
63
Things to do before starting
63
Procedure
64
Procedure
64
Protocol: RNA Cleanup
67
Protocol: RNA Cleanup
67
Determining the correct amount of starting material
67
Determining the correct amount of starting material
67
Important points before starting
67
Important points before starting
67
Things to do before starting
67
Things to do before starting
67
Procedure
67
Procedure
68
Procedure
68
Troubleshooting Guide
70
Troubleshooting Guide
70
Appendix A: General Remarks on Handling RNA
73
Appendix A: General Remarks on Handling RNA
73
Handling RNA
73
Handling RNA
73
General handling
73
General handling
73
Disposable plasticware
73
Disposable plasticware
73
Nondisposable plasticware
73
Glassware
73
Nondisposable plasticware
74
Nondisposable plasticware
74
Glassware
74
Electrophoresis tanks
74
Electrophoresis tanks
74
Solutions
74
Solutions
74
Appendix B: Storage, Quantification, and Determination of Quality of RNA
76
Appendix B: Storage, Quantification, and Determination of Quality of RNA
76
Storage of RNA
76
Storage of RNA
76
Quantification of RNA
76
Quantification of RNA
76
Spectrophotometric quantification of RNA
76
Spectrophotometric quantification of RNA
76
Purity of RNA
77
Purity of RNA
77
DNA contamination
77
DNA contamination
78
Integrity of RNA
79
Integrity of RNA
79
Appendix C: Formaldehyde Agarose Gel Electrophoresis
80
Appendix C: Formaldehyde Agarose Gel Electrophoresis
80
FA gel preparation
80
FA gel preparation
80
RNA sample preparation for FA gel electrophoresis
80
RNA sample preparation for FA gel electrophoresis
80
Gel running conditions
80
Composition of FA gel buffers
80
Gel running conditions
81
Gel running conditions
81
Composition of FA gel buffers
81
1x FA gel running buffer
81
1x FA gel running buffer
81
5x RNA loading buffer
81
5x RNA loading buffer
81
Appendix D: Optional On-Column DNase Digestion with the RNAse-Free DNase Set
82
Appendix D: Optional On-Column DNase Digestion with the RNAse-Free DNase Set
82
Important points before starting
82
Important points before starting
82
Things to do before starting
82
Things to do before starting
82
Procedure
83
Procedure
83
Appendix E: DNase Digestion of RNA before RNA Cleanup
85
Appendix E: DNase Digestion of RNA before RNA Cleanup
85
Important points before starting
85
Important points before starting
85
Things to do before starting
85
Things to do before starting
85
Procedure
85
Procedure
86
Procedure
86
Appendix F: Acetone Precipitation of Protein from Buffer RLT Lysates
87
Appendix F: Acetone Precipitation of Protein from Buffer RLT Lysates
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Equipment and reagents to be supplied by user26F
87
Important point before starting
87
Important point before starting
87
Procedure
87
Procedure
87
Appendix G: RT-PCR and Real-Time RT-PCR
89
Appendix G: RT-PCR and Real-Time RT-PCR
89
RT-PCR
89
RT-PCR
89
Real-time RT-PCR
89
Real-time RT-PCR
90
Real-time RT-PCR
90
Ordering Information
91
Ordering Information
91
Document Revision History
98
Document Revision History
98
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