Duotech srl Via Monte Spluga 31 - 20021 Baranzate (Milano)
tel. +39.0233106630 fax +39.0233106640
www.duotech.it [email protected]
*lv
. New Een€rotion polymÉrose urith supÈrior performf,ncÈ
r Novel buffer syst*m, with ultro-pure dNTPs ond ÀigCl,
. Robust ond high yield ocrass E full rongs of templotes
. Convenient oll-in-one moster mix
. Direct gel looding
The MyTaq* product range is a new generation of very high performance PCR products cjeveioped
l:y Eìoline. De*igned to deliver outstanding reeults on all ternplates, includìng cornplex genornic DNA
templates. MyTaq ìe baeed on the lateet technology ìn FCFI enzyme preparation, engineered to increase
tffinlty for DNA, reaulting ìn significant improvementa to yieid, senaitivity and epeed. The
enzyn-re is
supplied with an industry-ieading novel buffer system, speciiical{y f*rrnulateel end ualidated for the unique
propertiea o MyTaq, rnnkinrg it the perfect choice for all of your PCR assays.
g.Emic DilA{Al% GC Entsìt}
pollmsms from othÈr flpdieE lDr fiÈ mpllficElim of a 460bp lregmEnt of the hum f,Ue gÈnB, De§rÈasino ffiunts of hutM
ttrnplat€ {1p9, l0&ì8, i00ng, EOrE, 26ng@d 12,6ngitam 1-6 NpsÉtEtylln tls FCFI, ThE cvclrE re pÉrfbmad urdsrthB foilflinE
aondition§:.g6ic fJr 6min, fdbv#d by 3ù qrcles at S't tor 3Ds, 60.C brSB 6nd IZ'C ftr EOB. irErkÉr is FlypÈrLEdd$ I (M) {Cat No, Bt0_gg0!6),
WTq dÈtt ÈE
lighBr yi€ld and BÈNilivity N compsr€d !jlh all fN mmpÉflna prEdEte,
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[email protected]
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MyTaq - Full range of templates
MyTaq is a hiEh performance polymerase whieh exhibits rnore
robust ampliiication than other commonly used polyrnerases
(fiq. 1). MyTaq offers higher yields over a fult range of PCR
templates. nraking it the ideal choice for moet routine assays.
This new enzyme from Bioline is supplied with the MyTaq buffer
§ystem, a proprietary formulation containing ultra-pure dtrlTPs,
MgCl, and enhancers at optimal concentralionsl removing the
need for optimkation and giving euperior amplìfication"
MyTaq - For all applications
Thie new generation DNA polymerase from Bioline has been
validated with a full range of templates and is perfectly suited for
the follo'ruing applications :
e
.
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o
High-throughput PCH
Specific ampli{ication of complex templates
Bobuet ampli{ication of GC-rich sequencee
Routine PCR applications
TA cloning
MyTaq - For faater FGB reactions
The advanced forrnulation of MyTaq allows faeter FCR reactione
than other conventional polymeraees, thus reducing the overait
time from over an hour to leee than thir-ty minutee and most
importantly, without cornpromising PCR speci{icity or yield {fig. 2).
Reducing the reaction time ailov,rs greater lhroughput afld fast€r
screening.
Fig. ? Fest atrplitkation o, human gEmmic DfA {p€rfomìcd in 2L5 rinutes}
ComFaratire mFliisdian o, a 460bp fEgmst ol ttÉ huflan F]6 gme (61 % ccl
used io oompffi lolaq with a conwntiord fag ONA polymelffi. lh€ PCB
ws perbrmsd usim both ÉElmssusing d€c@ingamuntsof humm gercmic
DNA astemdaia(200nq, 66n9, 10n9, gng, 1n0,300p0. 10Op€ ild 30psl tseE
l-8 Espstivsly) ard undqr tlE icll@ing IEstsrcling conditiom: gEdc for 3min,
fdwEd bry 30 c!&ls st gs'C tur 15s. 60oC lor 15s ild 72"C ior 15§. Marksr i§
HypsLaddsr I [M) (Cat llr. BIO-33@5). MyTaq rsdilycoFeswith ttrb.rsction3
tim6, EEulting in highqryiÉld uithoul fis n6Èd for turllsopiimization.
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MyTaq - Oirect gel loeding
MyTaq is also supptied as MyTaq Red DNA Polymeraee, which
includes a 5x colored reaction buffer with an inert red dye.
Following PCB, samples can be loaded directly onto the agarose
gel without the need for a loading buffer, since the mix is of
sufficiently high density to sink to the bottom of the weli.
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MyTaq - Pramixee to simplify PCR eet-up
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MyTaq 2x Mix and MyTaq Red 2x Mix contain all the reagents
(inclucling stabilizers) necessary for eetting up a trouble-free
PCR reaclion. These novel mixes, supplied conveniently in one
tube, reduce the number o{ pipetting stepe and facilitate greater
eff
lì,:
iciency. thro ughput and reproducibi[ity.
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Tel +39.0233106630 fax +39.0233106640
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Stor4e and stability:
The MyTaq is shipped on Dry/Blue lce and can be stored ficr up to 12 rnorìth at -20'C, or up to
2 ì,veeks at +4"C. Repeated freeze^havv cydes should be avoided.
Shipping: On Dry/Blue
Exp. Date: See
lce
vial
Catalog numbers
BIO-21 105
:
Safety precautions:
Hannful if swallo,ed. lrritating to eyee, respiratory system and skin. Please refer to the material
500 Units
Batch No.: See vial
Blo-21'106: 2500 Units
Concentration:su/pl
BIO-21107: 5000 Units
safety data sheet for further information.
Unit definition:
One unit is defined as the amount of enzyme that incorporates l0nmoles of dNTPs into acidinsoluble fiorm in 30 minutes at 72"C.
This product insért is a declaration of analysis at the time of manufacture.
Research Use Only.
BIOLINE
srl "guo fata vocant'- www.duotech.it [email protected]
Description
MyTaqrM DNA Polymerase is a high performance PCR product that exhibits more robust amplification than other commonly used
polymerases, delivering very high yield over a wide range of PCR templates and making it the ideal choice for most routine assays. This
new enzyme preparation from Bioline is supplied with MyTaq Reaction Buffer system, an advaned formulation that savès time and
delivers superior results, containing dNTPs, MgCl2 and enhancers at optimal concentrations which eliminates the need for optimization.
Gomponents
lmpoÉant considerations and PCR optimization
The optimal conditions will vary from reaction to reaction and are
dependent on the templatelprimers used.
The
5x ltlyTaq Reacfon Buffer:
5x MyTaq Reaction Buffer
oompdses 5mM dNTPs, 15mM MgCla, Etabilizers and enhancers. The
concentration of each component has been extensively optimized,
reducing lhe need for further optimization. Additional PCR enhancers
such as HiSpec, PolyMate or Betaine etc. are not recommended.
14 x 1.5m1
Standard MyTaq Protocol
The following protocol is for a standard 50pl reaction and can be
used as a starting point for reaction optimization. All reactions
should be set-up on ice.
Primens: Fonitrard and reverse primers are generally used at the final
concentration of 0.24.6pM each- As a starting point we recommend,
using 0,4pM as a final concentration (i.e. 2Opmol of each primer per
50pl reaction volume). Too high a primer concentration can reduce
the specifici§ of priming, resulting in non-specific products.
designing primers we recommend using primerdesign
software such as Primer3 (htp://frodo.wi.mit.edu/primer3) or visual
OMPrM (http://dnasoftware.com) with mdnovalent and divalent cation
concentrations of 1OmM and 3mM respectively. Primers should have
\Men
PCR reaction set-up:
5x MyTaq Reaction Buffer
1Opl
Iemplate
rs required
Primers 20pM each
1pl
MyTaq DNA Polymerase
).25 - 11tl
ullater (ddHzO)
a melting temperature (Tm) of approximately 60'C
Template: The amount of template in the reaction depends mainly on
the type of DNA used. For templates with low structural complexity,
such as plasmid DNA, we recommend using 50pg-10n9 DNA per 50pl
reaction volume. For eukaryotic genomic DNA, we recommend a
starting amount of 200n9 DNA per 50pl reaction, this can be varied
between 5ng-500n9.
important to avoid using template
re-suspended in EDTA-containing solutions (e.9. TE buffer) since
EDTA chelates free Mg'".
lt ls
tp to 50pl
PCR cycling conditions:
lnitial Denaturation: An initial denaturation step of lmin at 95"C
Step
Temperature
Time
Cycles
lnitial denaturation
95"C
lmin
1
Denaturation
95"C
15s
Annealing
55"C
15s
Extension
72"C
10s
25-35
is
recommended for non-complex templates such as plasmid DNA or
cDNA. For more complex templates such as eukaryotic genomic
DNA, longer initial denaturation times of up to 3mins are required in
order to facilitate complete melting of the DNA.
Denaturation: Our protocol recommends a 15s cycling denaturation
step at 95"C which is also suited to GC-rich templates, however for
low GC content (4045o/ol templates, the denaturation time can be
decreased down to 5s.
and time; The optimal annealing
temperature is dependent upon the primer sequences and is usually 2
-5'C below the lower Tm of the pair. We recommend starting with a
55'C annealing temperature and, if necessary, to run a temperature
gradient to determine the optimal annealing temperature. Depending
on the reaction the annealing time can also be reduced to 5s.
Annealing temperature
* These steps may require optimization, please
if needed.
MT10-O9e
refer to the PCR optìmization section
Vllebsite: wìM,ì/.bioline.corn/ èmail: [email protected]
Extension temperature and time: The extension step should be
performed at 72"C. The extension time depends on the length of the
amplicon and the complexity of the template. \Mth low complexi§
template such as plasmid DNA, an extension time of "l0s is sufficient for
amplicons under 1kb or up to Skb. For amplification of fragments over 1kb
from high complexi§ template, such as eukaryotic genomic DNA, longer
extension times are recommended. ln order to find the fastest optimal
condition, we suggest incrementing the extension time successively up to
30s/kb.
Troubleshooting Guide
-
Check reaction set-up and volumes used
-
Check the aspect and the concentrations of all components as well as the storage
conditions. lf necessary test each component individually in controlled reactions
-
lncrease enzyme quantity to up to 2Ul50p1 reaction
Decrease the annealing temperature
Run a temperature gradient to determine the optimal annealing temperature
lncrease the extension time, especially if amplifying long target
lncrease the number of cycles
Cycling conditions not optimal
Decrease the number of cycles
Smearing
Annealing temperature too low
lncréase the annealing temperature
or
Decrease primer concentration
Non Specific
producb
-
Make sure all reactions are set-up on ice. Run reaction as quickly as possible
Replace each component in order to find the possible source of contamination
Set-up the PCR reaction and analyze the PCR produc* in separated areas.
Technical SuppoÉ
Associated Products
lf the troubleshooting guide does not solve the difficul§ you are
experiencing, please contact your local distributor or our Technical
Support with details
of
reac{ion setup, cycling conditions and
relevant data,
Email:
[email protected]
TRADEIIiARKS
1). Hyperladder and MyTaq are Trademarks of Bioline Ltd.
Bioline Ltd
UNITED KINGDOM
Bioline USA lnc.
USA
Bioline GmbH
GERMANY
Bioline (Aust) Pty. Ltd
AUSTRALIA
Tel: +44(0)20 8830 5300
Fax: +44 (O)2084522822
Tel: +1 508 880 8990
Fax +1 508 880 8993
Tel: +49(0)33 7168 1229
Fax +49 (0)337168 1244
Tel: +61 (0)2 9209 41 80
Fax +61 (0)2 9209 4763
DUOtgCh ttl Vi" Mont" Spluga 31 -20021 Baranzate (Ml)
tel. +39.02.33106630 fax +39.02.33't06640 www.duotech.it [email protected]
7!
Listino Prezzi MyTaq'* Bioline
Cat No:
q.tà
Descrizione
Prezzo €
BIO-21105
5(D units
MyTaq DNA Polymerase
€ 115
BIO-21106
25fi1 units
MyTaq DNA Polymerase
€ 380
Bto-21107
SfiX! units
MyTaq DNA Polymerase
€ 57s
Bro-21108
500 units
MyTaq Red DNA Polymerase
€ 115
Bro-21109
2500 units
MyTaq Red DNA Polymerase
€ 380
Blo-211X0
5ul0 units
MyTaq Red DNA Polymerase
€ 675
Blo-21111
250 units
MyTaq HS DNA Polymerase
€ 115
Bto-21112
lfiX) units
MyTaq H§ DNA Polymerase
€ 385
Bto-21113
25fi! units
MyTaq HS DNA Polymerase
€ 875
Bto-21114
250 units
MyTaq HS Red DNA Polymerase
€ 115
Bto-21115
10fi1 units
MyTaq HS Red DNA Polymerase
€ 400
Bto-21116
25fi) units
MyTaq llS Red DNA Polymerase
€ 87s
Blo-25041
2(X) Reactions
MyTaq Mix,2x
€ 100
Bto-2il42
10fi1 Reactions
MyTaq Mrx,2x
€ 430
81(>25043
2fi)
Bro-25044
€ 100
1000 Reactions
MyTaq Red Mix,2x
€ 430
Bto-25{r45
2OO Reactions
MyTaq HS Mif 2x
€ 1s5
B10-25046
10O0 Reactions
MyTaq H§ Mix,2x
€ 690
Br0-25@7
2fi)
Reactions
MyTaq HS Red Mix,2x
€ 155
Bto-2s048
l(XX) Reactions
MyTaq HS Red Mix,2x
€ 690
Duotech.20l0
MyTaq Red
Mir
2x
MyTaq a MyTaq
Reactions
HS comes with the dNTPs and MgCl2 already in the Reaction Buffer.