Thermo Fisher Scientific GeneJET Plant RNA Purification Kit User guide

Brand
Thermo Fisher Scientific
Model
GeneJET Plant RNA Purification Kit
Type
User guide
PRODUCT INFORMATION
Thermo Scientific
GeneJET Plant RNA
Purification Mini Kit
#K0801, #K0802
www.thermoscientific.com/onebio
#K0801, #K0802
 
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CERTIFICATE OF ANALYSIS
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78889:0)
8)
;7008)
070)
<07<0;=
088=
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B$!C#D3
7#"!3
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8?B:880.
9<09808--
2
COMPONENTS OF THE KIT
GeneJET Plant RNA
Purification Mini Kit
#K0
80
1
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#K0
802
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D
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STORAGE
5!"!/-%D)%F71
Note. Close the bag with GeneJET RNA Purification Columns tightly after each use!
DESCRIPTION
G !$!!!
$ #'!(#!#5HD
"!"$#(C$!
(CDCD$
!!!5)&'$##!(#
"'!C!$!$ #
"!'!$!'D7CD 7C
"$!D"!#"-##!(

PRINCIPLE
7#!"$$!#C"
$!$#!'!!!
!"!$
"!$#$0(#(!
" '!!'$!'
D'(!!5
3
Table 1.##!%&$(
Plant TissueYield, µg
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IMPORTANT NOTES
• !!!!(/.*D-&&I1Wash Buffer WB 1/!1!
Wash Buffer 2/!1J
#K0
80
1
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#K0
802
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Wash Buffer
WB 1Wash Buffer 2
Wash Buffer
WB 1Wash Buffer 2
7!'
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Ethanol (96
-
100%)
2.1
mL
3
0
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10.5 mL
100 mL
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!
• B#
!';/;CL&,*-1!!-&M);%&&M
Plant RNA Lysis Solution!
• 75Plant RNA Lysis Solution"D!(#
"#'$=3F7C"5!')%F7"
• E$('!$Plant RNA Lysis Solution and Wash Buffer WB 1 
/--9<098081!
!'5C!''!
• CC/-%D)%F71
4
ADDITIONAL MATERIALS AND EQUIPMENT REQUIRED
• !
• /.*D-&&I1
• ;CL&,*-
• -%$"
• $
• ;"$(
• E"+"%*
F
7
AVOIDING RIBONUCLEASE CONTAMINATION
#!$#!'"!$!!"#
C'$#"!#"#(7
"5!C#!$'
'EB)!!(!
J
• E$('!$$!C5
7$$( #
• ?C!"D
• ?$(!"
/C$1!'5
• 5$#!'$"
!#
RNA HANDLING AND STORAGE
• !''5$'
• !D)&F7D3&F79$"#C5
D3&F7
5
PROTOCOLS
$!C#D!#"!
!"!3
Before starting: Supplement the Plant RNA Lysis Solution with DTT (#R0861). Add 10 µL
of 2 M DTT to each 500 µL of Plant RNA Lysis Solution required for the extraction
procedure.
A. Plant RNA Purification Main Protocol
Step Procedure
1
%&&MPlant RNA Lysis Solution,!';C-%
$"/(!!1
E$D-&&$HN)&$
#H!
!$'$!J
1 !
-&&$ !$!$!$#'
!
"1 !$
-&&$($"!(
!"!!"!' !$
!!!#
0!#'!-%$"
$%&&M#A-&D)&$#
Note
• 

•  !
!
•
"#$%&'

2 0"=%*F77$%2)&C&&&
×
$/2-@C&&&1
3 7/#@%&D%%&M1!
$"/(!!1!!)%&M.*I"#$
4
!!"
7$--)C&&&×$/O--C&&&1;!'D
$!"!"
Note. '!"()'*
5
!!3&&PEBEB-/
"!!!EBEB-17$--)C&&&×$
/O--C&&&1;!'D$!"
)"
6
6
!!%&&PEB)/"
!!!EB)17$--)C&&&×$/O--C&&&1;!
'D$!"!"
7
*!D-!2)&C&&&
×
$
/2-@C&&&1
;!"$'D$!
D-%"
8
C!!%&PD'
"!$--)C&&&×$/O--C&&&1
Note+!,%- 
9
;!?!!#!'
D)&F7!
''5$'
Note../01#$%&'
7
B. RNA Purification from Lignified, Polyphenol-rich Plant Tissues
#'!#C$!!+#D"C
'$C!C'D!(/$C(1!'C#
'#(##!/A1
Step Procedure
1
0-%$"/(!!1!!%&&MPlant RNA Lysis
Solution!';/!"!%1!A)I/'+(1

!-&&$ !$$!
$!$!"!-%
'!!#-%$"
$%&&M#C!';!A
A-&D)&$#
2+3
 0
2 0"%%*F77$%2)&C&&&
×
$/2-@C&&&1
3 != %
C. RNA Purification from Soybean seeds
##"##'!
7!
Step Procedure
1
0-%$"/(!!1!!%&&MPlant RNA Lysis
Solution!';/!"!%1!7)

!-&&$ !$$!
$!$!"!-%
'!!#-%$"
$%&&M#C!';!7
A-&D)&$#
2+3
 0
2 !)%

8
ADDITIONAL PROTOCOLS
D. Genomic DNA Removal from RNA Preparations
1. Using DNase I, stand-alone product (#EN0521)
Step Procedure
1
!!D"J
 -P$
-&Q"'$7
)
 -P
;0CD/L&%)-1 -P/-1
ECD/L&%,-1 -&P
0"=3F7=&
Note. 34
/5)2%,6013 078
2
!!-P%&;!"*%F7-&(;0
!!; !#!#H!$$'!(
"$$
3 ?!!#!'
Note
• ;-;0-P$
• A!%&;"!$
!!&-P$+P
• E'5$'!!C("!
!$:'(C
-;+M$!O%;
2.Using Thermo Scientific RapiOut DNA Removal Kit (#K2981).
5!$!(($;!" 
(;0'D!";0CD
!#;($;0(
9!!C('''+"
9
TROUBLESHOOTING
Problem Possible cause and solution
Low RNA yield
Insufficient homogenization of plant material.
!'C$H
$#$!'!
Excess sample used during lysate preparation.
!$;-&&$

Ethanol was not added to the lysate.
'!!!#"#$

Ethanol was not mixed properly with the lysate.
!!#C"#($
$
Ethanol was not added to Wash Buffers.
'!!!EBEB-!EB)
"
9'EB=
DTT was not added to Plant RNA Lysis Solution.
;!"!!! #
!
Sample may be old or degraded
0"#$(
Purified RNA is
degraded
RNase contamination.
(!"C'$( !$(#
!(($C!" 
7$$( #?C!"
D!"!'5
'!$!
Inappropriate sample storage conditions.
9DH!$H !$
0!#$H!'!#

!!"!!#!'
!D)&F79$!$
/-1H$D3&F7!!
Inhibition of
downstream
enzymatic
reactions
Purified RNA contains residual ethanol.
0!'$
'EB)C#"!D
!!-!/2)&C&&&×$C
--C&&&1
Purified RNA contains residual salt.
?!EB'#'
'EBEB-!!
'$'EB)
10
Problem Possible cause and solution
Column clogging
Clarified supernatant contaminated with cell debris.
5#!#
#$$
To low temperature during centrifugation.
'!!$
)&D)%F7!$$
DNA contamination
DNA rich plant sample.
$;D!'('
!!""#(
$;!$';0/L&%)-1!
,
11
SAFETY INFORMATION
Plant RNA Lysis Solution
Xn:
:H!D!$"$JGuanidinium thiocyanate
Risk phrases
)) :''!
=*+=, 0$#!5
Safety phrases
)= ;"$++(+#
)* 0'#C!#'#'!5!
!(
=*+=3 E"($!$(
*& !"!!H!'
Wash Buffer WB 1
Xn:
:H!D!$"$JGuanidinium hydrochloride.
Risk phrases
)) :''!
=*+=, 0$#!5
Safety phrases
)= ;"$++(+#
)* 0'#C!#'#'!5!
!(
=*+=3 E"($!$(
*& !"!!H!'
PRODUCT USE LIMITATION
!!(!C!$!!!(#!#
!'!!$!$!(C"!

'''+"#;!
R)&-@90$(!!5#9
0!"!
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Thermo Fisher Scientific GeneJET Plant RNA Purification Kit User guide

Brand
Thermo Fisher Scientific
Model
GeneJET Plant RNA Purification Kit
Type
User guide
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