2. Thermo Fisher Scientific
  3. Maxima Hot Start Green PCR Master Mix, k1062

Thermo Fisher Scientific Maxima Hot Start Green PCR Master Mix, k1062 User guide

  • Hello! I'm a chat assistant that has read the user manual for the Maxima Hot Start Green 2X PCR Master Mix from Thermo Scientific. This document provides detailed information on using the master mix for PCR, including protocols, guidelines for primer design and preventing contamination and components of reaction mixture. The master mix features a ready-to-use format, hot-start polymerase and direct gel loading, ensuring high yield amplification. I am ready to answer your questions about this product.
  • What is the composition of the master mix?
    What is the recommended temperature for the extension step?
    What is the recommended concentration range of the PCR primers?
PRODUCT INFORMATION
Thermo Scientific
Maxima Hot Start Green PCR
Master Mix (2X)
#K1062

Lot: Expiry Date:
Store at -20°C
}
Ordering Information
Component
#
K10
6
1

#
K10
6
2




 
 !
"

 
www.thermoscientific.com/fermentas
Description
#$!!
%&""  !'
(Taq)*+&,-%
$. '/%%*##$
 ,,%0$0!1'%&%
%&'$0%!%'
,% !'#$%&$%
0$,!%!,.
0$%0,,! !$ !
 !)*+2 !''%!
%'#$ 
Taq)*+&3!,.
$'$&%,!' ,41.
'!)*+
5,,!$2 ,% !&.&
..! !!!0
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8
Applications
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• #"
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Maxima Hot Start Green PCR Master Mix (2X)
composition
Taq)*+,& ,,%
$. 9:%+#9:%#
9:%#9:%##%9'/#$
 ,,%0$%&
'%0%&%!%''
'
PROTOCOL
 &8%.&! '
$0'
 +%%$0'!,!$
!, ;
Maxima
Hot Start Green PCR
Master Mix (2X) 
Forward primer

"

Reverse primer

"

Template DNA
,
'
"
'
Water, nuclease
-
free
6<

Total volume

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9
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$%%8&$! 0$

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!&!'!% %.0;
Step
Temperature,
°C Time
Number
of
cycles
3% =
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) 
>
4
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+'
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4
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@
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5?
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GUIDELINES FOR PREVENTING CONTAMINATION
OF PCR REACTION
) '$!,,)*+
'%#$! .18%
!0$$,%,!$
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!%0$1!
0;
• ,& )*+, ,$ 
,$!&!'%&-,% !
,
•  , 0!.
2 ,,%0$CD,
• $'8)*+,!%! ,
• C'!%%!%C
,8%,!,,,,,0$
,,)*+,%,
 ,
• +0&,E,!F*#!
!$!1!
GUIDELINES FOR PRIMER DESIGN
C$#$!!?80,%'
0000!=800$
'!%,%'
 %.0;
• ,'&"4 !%'
• G,!$,9"AH3%&
% !%$ %.%. % &
'$,
• +8%,!'$$ !%
$4I"%0$1",!!,'
• 3,.$,$ %0$
$4I"%
• +8%"!,&,'
!,.0$,%%!
,,,8$,%,
%-
• $!1,. %%!,&
.0,%,)*+
• $%''%',,!
4!8% !%$4I"%
• $% !'!-&,
$.E8'!!&!$
'F!%
000$!!!=%$
 ..2 %!!8'
Estimation of primer melting temperature
5,!'$ !%$
,,', #!.!! %
'$0'2 ;
#J9//+/#
0$+#,$ .,!8 !%
$,
3$,!$ !%0
!% ',!-%!, ,''
?80K000!=80!! 
!%L!.!
!!!
COMPONENTS OF THE REACTION MIXTURE
Template DNA
G, ,)*+!
8 "'.$,%%,$')*+
%"''!)*+'$ 
,!$1'",!!
,% !B0 ,% !$
!! !&$,!
+ )*+, !$%!. %
,,$,'!)*+ !7
67#$!!M?#%
,,767=4#! !'
%)*+, !!$,$?)#+%
,7!$.)*+,&?$
,!,%,%0$$)*+,0$
@H$ &8!!)*+
,
Primers
#$!%%!!'$
,"?!8,!!
!$,..&,'%'
",!!,% !
5%',0!%$'$,
!!$'4":
(continued on reverse page)
CYCLING PARAMETERS
Initial DNA denaturation and enzyme activation
3!,&% $,)*+
$.''$  ! -
$,% '$,!!&!
Taq)*+,&!8
, % '$! ,%
!8%9% '$% =-&
!8,
Denaturation
+)*+% 4!%,!&!
>N& !5"!$)*+,
$,!.,'%4"9
Primer annealing
#$', $ %.N0$$
', #$,+'
4!%& !3",!!
,% !,,$', $ %.
,-%,0"N!
Extension
#$,, 
Taq)*+,&@"@N#$!%%
,=1.@N
Number of cycles
3$!,$,,$
!. 9!&!2 %5$'$
, "4!&! !
Final extension
+$!&!!%%! .$
 @N%%""
&,.!,!,% !3$
,% !0.!%#+8!!
'#$!!3#+!'7
674$,&.,'%
4 $!,4I"%+'$
,% !3$,% !0. %!' '
#$!!M?#'7674
$,!.%
Troubleshooting
5 .$',8
000$!!!=
CERTIFICATE OF ANALYSIS
Endodeoxyribonuclease Assay
*%!.!8!8&!%!! 
)*+!1%)*+0.8%! .

(

0$',C>)*+! 
9$ 4@N
Ribonuclease Assay
B$H$%!8&0%
!$!!!%" .!! .

(

0$'O
4
P"*+! 
9$ 4@N
Labeled Oligonucleotide (LO) Assay
*%!.%'%'"%%%
% ."%%.%' !%0.8%
! .0$9$ 4@N
Functional Assays
• ;,!49.,)*+'$
'!,&+G?','$ '!
)*+
• #";,!9>A.,)*+'
!)*+&$-% '#$!!8+%
 8#!,'*+
!,
Quality authorized by: M 'Q1
NOTICE TO PURCHASER: LIMITED LICENSE
C$,% !!8%.&$0'C,
%!,%',! %$C;A@A@@
!4&%@@4<!%A&#$, !$
$,% !! %%". & 
%$'',! '&$ ,% !
$, !$I0!$*'$ %&$,
!%'$,!!8!&1%! %'
0$ ,'$ , !$R!8
$!!!%!8&%,&.&,!
.&,,#$,% !!$ &)'!  %
!$,2 ,!!$5 $
, !$'!&..%.&!!'$)!
B!'+,,%S&<B!)85&
PRODUCT USE LIMITATION
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$ .
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