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

Type
User guide
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,,!$2 ,% !&.&
..! !!!0
!% '$!
67
8
Applications
'$$ '$, 
#"
'$&,!!,!!,'!
%!)*+,
+,!0!,&)*+'
,% !#+!'
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

4 &8$,%,%0
9
$ '$!&!$%!
$%%8&$! 0$

  '$!%%$
!&!'!% %.0;
Step
Temperature,
°C Time
Number
of
cycles
3% =
-&!8 > 9 
) 
>
4
"9
+'
#
"
4
?
@
=1.
5?
@
"

A B%" %!&'
GUIDELINES FOR PREVENTING CONTAMINATION
OF PCR REACTION
) '$!,,)*+
'%#$! .18%
!0$$,%,!$
&.,$.&8
!%0$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 $
, !$'!&..%.&!!'$)!
B!'+,,%S&<B!)85&
PRODUCT USE LIMITATION
#$,% !%8,%%'%%%! 8&for research
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Thermo Fisher Scientific Maxima Hot Start Green PCR Master Mix, k1062 User guide

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