2. Thermo Fisher Scientific
  3. GeneJET RNA Purification Kit

Thermo Fisher Scientific GeneJET RNA Purification Kit User guide

  • Hello! I'm your chat assistant, and I've reviewed the user guide for the Thermo Scientific GeneJET RNA Purification Kit. This kit is designed for quick and efficient RNA isolation using a silica membrane spin column method. The kit can be applied to a wide range of sample types and includes protocols for genomic DNA removal and RNA cleanup. I'm here to help with any questions you might have regarding the kit and its operation. Feel free to ask me about the procedures, components, or troubleshooting.
  • What is the principle of GeneJET RNA Purification Kit?
    What kind of samples can be used with this kit?
    How should I store the components of the kit?
PRODUCT INFORMATION
Thermo Scientific
GeneJET RNA Purification Kit
#K0731, #K0732

www.thermoscientific.com/onebio
#K0731, #K0732


CERTIFICATE OF ANALYSIS
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
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Quality authorized by: ";(:
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1=%<$##<9
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$6C1#6<=$1<#/3
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2
COMPONENTS OF THE KIT
GeneJET RNA Purification Kit 50 preps
E&)39/
250 preps
E&)39
%& +))F A
×
+))F
B
D)

))

GB/  
D)

))

GB   9 /))
G4 (
7

9)

/A

#$% 8(
7
(*
8((  A) A)
8(( 


A)
A)
8(( /A

A)
A)
STORAGE
%&(($(
7)H8< :(/A7AH8
Note. Close the bag with GeneJET RNA Purification Columns tightly after each use!
DESCRIPTION
!"I#$% &(  (#$
( ( ((44( ((4 4 
:(J( 7 ( 5 (4
(  (4( (  (7
( 
#$( (()) ( (*!"#$% 
&/A(7'(#$ *
*(  (7%847'%84#(#$7
(See(/ ((#$(5
PRINCIPLE
((JB4*   4
 ( ( #$#/(
*((  (  (( 
#$( *(( (
'(4 5(5* (*
*%#$((*  * (7
*
3
Table 1. ((#$(5 
Source Quantity Yield, µg
=
)
/)
7
/A
=
 (
9)
-
7
/)
=( 9) A79)
=:
9)
A
7
9)
=(5
9)
+)
7
+A
=( A /)7/A
Bacillus pumilis (( /
×
/)
0
 (( /A7)
Escherichia coli (( /
×
/)
0
 (( A79)
> (( A
×
/)
+
 (( 9A7D)
": (( A
×
/)
+
 (( D)7A)
83 (( /
×
/)
+
 (( )7A
Saccharomyces cerevisiae (( D
×
/)
-
 (( /A)7/+)
IMPORTANT NOTES
Buffer preparation and handling
$ 5((0+7/))KWash Buffer 1  
Wash Buffer 2  L
50 preps
E&)39/
250 preps
E&)39
Wash Buffer
1
Wash Buffer
2
Wash Buffer
1
Was
h Buffer
2
8 * D) 9 )) /))
Ethanol (96
-
100%)
10
mL
3
9
mL
50
mL
170
mL
(5(
A)

+

A)

3)

$(4:  :(M 5 
(
• B #$ 4('Lysis Buffer
*N7 ($)F/D9=N7 ()F=
 /5(Lysis Buffer
• 8 :Lysis Buffer(  7(5 
*(93H84 ( :*AH8
• G(5*(Lysis Buffer and Wash Buffer 1 ( 
see /3$6C1#6<=$1<#(  *:4(
*((*
4
Avoiding ribonuclease contamination
#$(*( #$ 
#$4* (( (58
: #$#$4 (( (**
GB(
!( 5# L
• G(5*(#$(4:  
#8(5'(
• @(4(#7
• @5# 7(
4 *: 
• &((: ((*$ (((
Starting material handling and storage
• G#$(4( ( (
5% (J' :((
• G(  ( 54(*
( (( ('(#$
• 1#$( (( 4J(('
73)H86J((((**
(*
• $( ( (( ((73)H8('>*54
#$  ((J (4((5( 

• B(( (( #$ ( ((( *
((DH8(J((
5
ADDITIONAL MATERIALS AND EQUIPMENT REQUIRED
• %(4#7
• O
• =  
• =  
• ((5
• '(J L
o =(
o 7J
o B((
• /D9=N7 (=
• (0+7/))K
Buffers
• 6( ( (((L
%B/93=#8(43=&8(4/)=#
>%<
D
4=&>
%<
D
4>3D
• 67575 (L
/)=>8(4>-)4/=$ (J)D,(
 
• 6(L
C(/=(4)/=$4>3D$)/K7N7 (
A)( J()P
• 6%&(L
/)=>8(4>-)4/=$
6
RNA PURIFICATION PROTOCOLS
% (#$ ( ( ((44 4(4
  +7/9
A. Mammalian Tissue and Insect Total RNA Purification Protocol
Before starting:
• ('B*N7 ($)F
/D9=N7 (= /5(B'
• %'%&(L(/)F%& (
A0)F/)=>8(4>-)4/=$
Step
Procedure
1
G9)J
4:

(((*L
 (
%( 9)/)( ('
(*( 
*(/A   ( 
9))QB(*N7 (O/)
(
Note
•  *B' :((
5***B (#$
•  ( (#$ (

• $((((*B(
(*((  #$
>J(7J(
()7(#75(
 J7J
%( 9)/)( (
J5(J 9))QB(*
N7 ((( 5(
7J)7D)(
2
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 (%&(
A0)FO( /A7AH8/)
3
8
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R/)(
/)
(2/)(./)))×
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7
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Procedure
5
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(( 8 (/./)))× (*7
(   ( : (( 
(((( (
  ((  (*7(%( 
!"#$% 8(* ((  (
Note. Close the bag with GeneJET RNA Purification Columns tightly after each
use!
6
$3))QGB/(*(4see9!"
#$% 8( /./)))× 
(*7(   ( : (( 
7
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#$% 8( /./)))× (*7
(   ( : (( 
8
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!"#$% 8( 
./)))×
Optional1((  (4 (( 
7 (/
  ((  (*7(
!"#$% 8((/A#7
  
 (
9
$
/))Q
G4 (
7
 ( !"#$
% 8(8/./)))×(#$
Note
• =0)K#$((6#$
((*(/))QG4 (7
• 1  #$'#$(((
(4RA5(G4 (7
  
A)Q
 (%(*((5(
((((('(#$
10
   (@#$*( 
#$7)H873)H8(
8
B. Mammalian Cultured Cells Total RNA Purification Protocol
Before starting:
• ('B*N7 ($)F
/D9=N7 (= /5(B'
Step
Procedure
1

 ((
%((/×/)
3
 ((  A
A)×  (( *%B5
(*  
$ ((
5* ((A×/)
+
 (( ((
 *%B5(5 %B 
(( (( 5(%B
J ((   (((
 AA)× 
Note
%(( ((  (#$(
7
3)H8(
2

 (( (( //
+))Q(B
(*N7 (O/)(
Note
• 1B 5 4
J('>J(9)
7J(()7
(
#75(1 (J*(((
  #$(
• 1 ((5(4 A
/D)))×*#7  
 (
3
$9+)Q
(0+
7
/))
K
(

4
3))Q(!"#$% 8(
 (( 8 (/./)))× 
(*7(   ( : (( 
(((( (
  ((  (*7(%(
!"#$% 8(* ((  (
Note. Close the bag with GeneJET RNA Purification
Columns tightly after each
use!
5
$3))QGB/(*(4see9!"
#$% 8( /./)))× (*7
(   ( : (( 
9
Step
Procedure
6
$+))QGB(*(4see9!"
#$% 8( /./)))× (*7
(   ( : (( 
7
$A)Q
GB!"#$% 8(
./)))×
Optional1((  (4 (( 
7 (/
  ((  (*7(
!"#$% 8((/A#7  
 (
8
$/))QG4 (7 ( !"#$
% 8(8/./)))×(#$
Note
• =0)K#$((6#$
(4(*(/))QG4 (7
• 1  #$'#$(((
(44S/×/)
+
 ((45(G4 (7

 A)Q (%(*
((5(((((('(#$
9
 
  (@#$*( 
#$7)H873)H8(
10
C. Human Blood Cells Total RNA Purification Protocol
Before starting:
• ('B*N7 ($)F
/D9=N7 (= /5(B'
• B(( (( #$ ( ((( *
((DH8(J((
Step
Procedure
1
8(( ( (( )A*((D))
×
A
DT8B( ((*(((((+)73)K((
5(5 ((((*
2
((+
))Q

B(*
N7 (O(
3 $DA)Q(0+7/))K
4

3))
Q
(!"#$% 8(
 (( 8 (/./)))× (*7
(   ( : (( 
(((( (
  ((  (*7(
%( 
!"#$% 8(* ((  (
Note. Close the bag with GeneJET RNA Purification
Columns tightly after each
use!
5
$3))QGB/(*(4see9!"
#$% 8( /./)))× (*7
(   ( : (( 
6
$+))QGB(*(4see9!"
#$% 8( /./)))× (*7
(   ( : (( 
7
$A)QGB!"#$% 8(
./)))×
Optional1((  (4 (( 
7 (/
  ((  (*7(
!"#$% 8((/A#7  
 (
8
$A)Q
G4 (
7
 ( !"#$
% 8(8/./)))×(#$
9
   (@#$*( 
#$7)H873)H8(
11
D. Bacteria Total RNA Purification Protocol
Important Note
6#$(  (((5(*
<
+))
U)A7/Do not use an overnight culture for RNA isolation.
Before starting:
• ('/)=>8(4>-)4/=$*
(J ((  )D,
• ('B*N7 ($)F
/D9=N7 (= /5(B'
Step Procedure
1
/A!75!75 ( (/
×
/)
0
((/A  8((  (( 
./)))8((54(5(((
2 ((/))F((*
(J)D,(  155(
3
1  ((A
/A
7
A
8
4
$9))Q

B(*
N
7
 (
=
(5/A(
5
$/-)Q
(0+
7
/))K

6

3))
Q
(!"#$% 8(
 (( 8 (/./)))× (*7
(   ( : (( 
(((( (
  ((  (*7(%( 
!"#$% 8(* ((  (
Note. Close the bag with GeneJET RNA Purification Columns tightly after each
use!
7
$3))Q
GB/(*(4
9
!
"
#$% 8( /./)))× (*7
(   ( : (( 
8
$+))Q
GB(*(4
9
!"
#$% 8( /./)))× (*7
(   ( : (( 
9
$A)Q
GB!"#$% 8(
./)))×
Optional1((  (4 (( 
7 (/
  ((  (*7(
!"#$% 8((/A#7  
 (
12
(continued)
Step
Procedure
10
$/))QG4 (7 ( !"#$
% 8(8/./)))×(#$
Note
• =0)K#$((6#$
((*(/))QG4 (7
• 1  #$'#$(((
(4S/×/)
+
 ((5(G4 (7 
 A)Q (%(*((
5(((((('(#$
11
   (@#$*( 
#$7)H873)H8(
E. Yeast Total RNA Purification Protocol
Important Note
6#$( (((5(*
<
+))
U)A7/Do not use an overnight culture for RNA isolation. 6 ((
J ( (*((5 ((
Before starting:
• %C(L/=(4)/=$4>3D"
)/Kβ7 (A)( J()
• ('B*N7 ($)F
/D9=N7 (= /5(B'
• %'%&(L(/)F%& (
A0)F/)=>8(4>-)4/=$
Step
Procedure
1
 (( (D
×
/)
-
 ((/A  
58((  (( ./)))× 

2

 ((((/))
F

C(
3
1 9)H89)
4
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8
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Note. Close the bag with GeneJET RNA Purification Columns tightly after each
use!
9
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10
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11
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!"
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./)))×
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#$
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Note
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13
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ADDITIONAL PROTOCOLS
Genomic DNA Removal from RNA Preparations
A. Using DNase I, stand-alone product (#EN0521)
Step
Procedure
1
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7
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 /Q
#14#7E#)A/ /Q/
%87GE)+)/ /)Q
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Note.    :#1E<)9-/4 ((/,Q4 
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2
$/QA)=$ +AH8/) 5#1
$$'#$(J*5( 
  (
$(5(4#$!"#$% &see/A#$
8(% ((, ( 
3
@#$*( 
Note
• /#1/Q#$
• O( A)=$(  ((
#$ (  #$)/Q,Q
B.Using Thermo Scientific RapiOut DNA Removal Kit (#K2981).
: 55(#$#$('5(
#1(*7 &  #14#7
#5( #15(
6(4(5***   ,
15
RNA Cleanup Protocol
!"I#$% &  (/))µ#$ (#1
J  
Step
Procedure
1
$P5( 
/))
F
*
G4 (
7

 ($9))FBwithoutN7 (=
(5
2 $/-)Q(0+7/))
%

3

#$% 8( (( 
8 (/./)))×  (( 
(*7(%( !"#$% 8(
* ((  (
Note. Close the bag with GeneJET RNA Purification Columns tightly after each
use!
4
$3))QGB/(*(4see9!"
#$% 8( /./)))× (*7
(   ( : (( 
5
$+))QGB(*(4see9!"
#$% 8( /./)))× (*7
(   ( : (( 
6
$A)Q
G
B!"
#$% 8( 
./)))×
Optional1((  (4 (( 
7 (/
  ((  (*7(
!"#$% 8((/A#7
  
 (
7
$A)Q
G4 (
7
 (
 !"
#$
% 8(8/./)))×(#$
Note. 6#$((*(A)Q
G4 (7
8
   (@#$*( 
#$7)H873)H8(
16
TROUBLESHOOTING
Problem Possible cause and solution
Low RNA yield
Too much starting material was used for lysate preparation.
 ( ((
 ( (
Starting material was not completely disrupted.
 (1 
Ethanol was not added to the lysate.
=:(*(((
  (
Ethanol was not added to Wash Buffers 1 and 2.
=:(*GB/
6((* GB9
Degraded RNA
Inappropriate handling of starting material.
G#$(4( ( (
5% (J' :((
J(J('( (( 
73)H8*((5(
B
RNase contamination.
5# 4*(5 ((  
(5'(@(4(#75#
7(*:
DNA
contamination
#$*#1E
#)A/

1 5#1
  (4(, ( 
7#$((*#$8(% (see/A
Column
clogging
Too much starting material was used for lysate preparation.
 ( ((
 ( (
Starting material was not completely disrupted.
 ( 
Tissue debris was not removed before applying lysate on a column
=(1 (#$% % (
8J ((5
(( (
Enzymatic
reactions not
running well
Ethanol was carried over into the eluted RNA.
1((  (**G
B4 (( 7 ((/
./)))×
Salt was carried over into the eluted RNA.
@GB$(**(*
GB/ **GB
17
SAFETY INFORMATION
Lysis Solution
>J7 ((LGuanidinium thiocyanate
Xn>(
Risk phrases
 >(*((*
9+,9- 1:
Safety phrases
9 ,,5,
+ 1   *4(*(*: (5 
9+,93 G( 5 ((5
+) ( J*
Proteinase K
>J7 ((LProteinase, Tritirachium album serine
Xn >(
Risk phrases
D = J(
Safety phrases
9 ,,5,
9+ G( 5 (
DA 1  (*((4: (5 (*((*
(
+) ( J*
References
/ 8 J:4%#  /0-3(7#$(  7
(7 ( $(B /+L/A+7/A0
 B448"$(4===(48"4%=G((4"5#/00)
(  (  "8(= (-LD0AWA)9
PRODUCT USE LIMITATION
 5(4( (5( in vitro ( 
* 5(4(
(
%(***   ,=( 
X)/D6  1 $((5$((:6
  1 
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