GoTaq DNA Polymerase

PCR
Certificate ot Analysis
REF M3001 LOT 0000447667
GoTago DNA Polymerase: 2023-03-0
Supplied With: Conc 5u/pt Dispensed Lot# 0004373 16
5X Green 5X Colorless
100u
Cat. GoTaq Bulfer GoTag Buffer
Size
M3001 1mi (M791A) 1ml (M792A) For Laboralory Use
100units
M3005 4x 1ml (M791A) 4x 1ml (M792A) Country of Ortgin: USA
M3008 500 units 4 x 5ml (M7921)
COporton
wo Wnede f e Nrad

2.500 units 4x5ml (M7911)

M3008-C 500 units
EEL
ouo
a proprietary formulalion containing HEPE
eSEription: GoTaq® DNA Polymeraseab) is a Ra DNA polymerase supplied inand M3008 are supplied wilh 5x Green
ADM3001 00004476676
designed for amotification. Cat.# M3001, M3005
Sand SX DuTers enhanced
Colorless GoTags Reaction Bulfers,TheSX Gieen Gotag Reaction Bulfer contains wo dyes (bue and yel0wunat
is used when direct 1uoresCence or
aheh g eEcropnoresis to monitor migralion progress. The colorless buffer
of the amplified DNA from the PCR. Both bufers contain MgClh
of 7
concenttation of
al a concentration
7.5mM
Without prior puritication
s for a final
concentration 15mM in the 1X reaction.
of
Biological Source: The enzyme is derived trom bacleria.
Enzyme Concentration: 5u/
XGreen GoTaq® Reaction Buffer (Part# M791A, M791B): Proprietary fomulation supplied at phH 8.5 containing
UE and yellow dyes. In a 1% agarose gel, the blue dye migrates at the same rate as 3-5kb DNA fragments, and the yeliow
oye migrales at a rate faster than primers (<50bp). Green GoTaq Reaction Buffer also increases the density of the Sample,
SD ft will sink into the well of the agarose gel, allowing reactions to be loaded directly onto gels without loading dye. This
butler contains 7.5mM magnesium. Vortex thoroughly after thawing and prior to use.

5X Colorless GoTaq Reaction Buffer (Part# M792A, M792B): Proprietary formulation supplied at pH 8.5. This
butfer contains 7.5mM magnesium. Vortex thoroughly alfter thawing and prior to use.
Promega
Storage Conditions: See the Product information Label for storage recommendations. Avoid exposure to frequent Promega Corporation
temperature changes. See the expiration datle on the Product Intormation label. 2800 Woods Hollow Road
Madison, Wi 53711-5399 USA
Unit Definition: One unit is defined as the amount required to catalyze the incorporation of
ofenzyme 10 nanomoles of Telephone 608-274-4330
dNTPs into acid-insoluble material in 30 minutes at 74°C. The reaction conditions are specified below under Siandard DNA Toll Free 800-356-9526
Polymerase Assay Conditions. 608-277-2516
rax
Internet www.promega.com

Quality Control Assays

Activity Assays PRODUCT USE LIMITATIONS, WARRANTY DISCLAIMER
Promega manufactures products for a nurmber of
Functional Assay: GoTaq DNAPolymerase testedfor
is performance in Using
the polymerasechain reaction (PCR) intended uses. Please referto the product label for the
125 units ofenzyme to amplity a 360bp region of the a-1-antitrypsin gene from 100 molecules (0.36ng) of human genome intended use statemens tor Specin po h

DNA. The resulting PCR product is visualized as asinge band on an ethidium bromide-stained agarose gel. ru Su raant direct human contact.
all Promega producis to prevent direct human contact
Standard DNA Polymerase Assay Conditions (Not PCR Conditions): The polymerase activity is assayed in 50mM
Tris-HCi (pH 9.0); 50mM NaCl, 5mM MgCl,, 200uM each of dATP, dGTP, dCTP, dTTP (a mix of unlabeled and [9H]dTTP); Each Promega product is shipped with documentation
stating specitications and othertechnical information.
Promega products are warranted to meet or exceed the
10ug activated caif thymus DNA; 0.1mg/ml BSA in a final volume of 50ul. stated specifications. Promegas sole obligation and the
customers sole remedy is limited to replacement o
Contaminant Assays producis Iree or charge in ine event producis tal to
pertorm as warranted. Promega makes no oner war
Reaction Buifer is tested to ensure that it
5X Green GoTaq° Reaction Buffer Migration Pattern: The 5X Green GoTag® ranty of any kind whatsoever, and SPECIFICALLY DIS-
loading dye agarose gel electrophoresis. 1kb DNA Ladder (Cat.#
for CLAIMS AND EXCLUDES ALL OTHER WARRANTIES
does not interfere with DNA migration when it is used as a
OF ANY KINO OR NATURE WHATSOEVER, DIRECTLY
either 1X Green GoTag Reaction Bufer or the Blue/0range Loading Dye supplied with the markers and OR INDIRECTLY, EXPRESS OR IMPLIED, INCLUDING.
G5711) is mixed with WITHOUTLIMITATION, AS TO THE SUITABILITY.
and ethidium bromide slaining, the migration distance and pattern of the
loaded on a 1% agarose gel. Following electrophoresis PRODUCTIVITY, DURABILITY, FITNESS FOR A PAR
bands in both lanes are identical. TICULARPURPOSE OR USE, MERCHANTABILIY,
CONDITION, OR ANY OTHER MATTER WITH

GoTaqe DNA Polymerase is determined to be >90% pure judged by SDS-polyacrylamide gel

as RESPECT TO PROMEGA PRODUCTS. In no event shall
Enzyme Physical Purity: Promega be lable tor claims tor any otner damages,
whether direct, incidental, foreseeable, consequential,
electrophoresis with Coomassie® blue staining. or special (including but not liited to loss of use, rev-
exonuclease activity detecled.
Nuclease Assays: No contaminating endonuclease or enue or prolit), whether based upon warranty, contract,
tort(inctudng negligernce) or strict habiity ariSingin
Connection witn the sale or the tailure ol Promega prod-
ucis to pertorm in accordance witn the stated speciica
PCR Satisfaction Guarantee Ions.
and reagents are proven in PCR to ensure relable, high-pertormance results.
Promega PCR Systems, enzymes ol Technical Support scientists.
Your success is importan! to us. dur products are backed by a worldwide team
Please contact them for- applicatione or technical agsiclance, lf you are not complotaly.satisfied with any 2002-2018 Promega Corporation. All Rights
Il send
a replacement or refund your account. Heserved

PCR Promega PCR product we

That's Our PCR Guarantee GoTaq and Wizard are registered trademarks of
PromegaCorporation.
date and have been stored and used in accordance with product literature. See
Product must be within expiration Coomassie is a registered trademark of Imperial
AR Promega Product Insert ior specific lests performed. Chemical Industries, Ltd.
Products may be covered by pending orissued
patents or may have certain limitations. Please visit
La-Roche or Applera. This Our Web site for more information
the US for basic
PCR
Is OUI any valid U palents assSigned to Hofiman
wIthout any license or royalty fees.
aUse of this product in agriosuc appications All prices and specifications are subject to change
basic PCR in researcn, u i a
product can be used for 2,335, 153, Chinese Pat. No. ZL9980886 1.7. Honoorig Kan
Kong without prior notice.
Australian Pat. N. /i dndolan PatL No.
Pat No. 1088060 and other patents pending,
(0)IU.S. Pat. No. 6,242,235, No. 3b/31/0, EuOpEdn Product claims are subject to change. Please contact
Japanese Pat. Promega Technical Services or access the Promega
Pat. No. HK 1040262,
online catalog for the mosi up-t0-daie intormation on
Promega products.

Part# 9PIM300
Printed in USA. Revised 4/18.

Signed by: R. Wheeler, Quality Assurance

Promega Usage Information
1. Standard Application B. Bulfer Choice
We recommend using the 5X Green GoTaqo Reaclion Bufferinany amplfication reaction
Reagents to Be Supplied by the User
that will be visualized by agarose gel clectrophoresis followed by etridium bromide
PCR Nucleotide Mix(Cat# C1141, upstream primer staining. The 5X Green GoTaq Reaction Butfer is not recommended for any downstream
145) downstream primer applications using absorbance or tluorescence excitation, as the yellow and blue dyes in
Nuclease-Free Water (Cat # P1193) the reaction buffer may interfere with these applications. The dyes absorb at 225-300nm,
template DNA making standard Azgo readings to determine DNA Concentration unreliable. Also, the dyes
1. In a sterile, nuclease-free microcentrituge tube, combine the following on ice: have excitation peaks at 488nm and al 600-700nm that correspond to the excitation
Component Final Volume Final Concentration wavelengths commonly used in fluorescence detection instrumentation. However, for
some instrumentation, such as a fluorescent gel scanner that uses a488nm excitation
5X Green or Colorless
Go Taq Reaction Buffer 10pl 1X (1.5mM MgCl, wavelength, there will be minimal interference because it is the yellow dye that absorbs
PCR Nucleotide Mix, 10mM each this wavelength. Gels scanned by this method will have a light grey dye front below the
1 0.2mM each dNTP
upstream primer Xul 0.1-1.0uIM primers corresponding to the yellow dye front. The Green and Colorless Go Taq® Reaction
downstream primer Buffers give approximately equivalent amplification yields. Balanced amplifications
Yul 0.1-1.0pM
GoTaq DNA Polymerase (5u/ul) between the two bufers may require further optimization.
0.25pl 1.25u
template DNA
<0.5ug/50ul For reactions going directly from thermal cycler to an application using absorbance or
Nuclease-Free Water to 50pl fluorescence, the 5X Colorless GoTaq Reaction Bufer is recommended. If both agarose
Completely thaw and thoroughly vortex the bufter prior to use.
More MgCi, can be added to the reaction using 25mM gel analysis and further downstream applications involving absorbance or fluorescence
MgCl, Solution (Cat.# A3511). will be used, the two dyes can be remøved from the Green GoTaq reactions using
2 If using a thermal cycler without a heated lid, overlay the reaction mix with 1-2 drops standard PCR clean-up systems like the Wizard® Sv Gel and PCR Clean-Up System
(approximately 50pl) of mineral oil to prevent evaporation during thermal cycling. (Cat. A9281) or Wizard® PCR Preps DNA Puriication System (Cat.# A2180).
Centrifuge the reactions in a microcentrifuge for 5 seconds.
Both reaction bufters are compatible with common PCR additives such as DMS0 and
3. Pertorm PCR using your standard parameters. An example profile is given in Table 1. betaine. These aditives do not change the color of the Green GoTaq® Reaction Buffer o
For the cycling protocol, we recommend the following: affect dye migration.
a. Reactions are placed in a thermal cycler that has been preheated to 95°C.
C. Primer Design
b. The thermal cycling protocol has an initial denaturation step where samples
PCR primers generally range in length from 15 to 30 bases and are designed to flank the
are heated at 95°C for 2 minutes to ensure that the target DNA is completely
region of interest. Primers should contain 40-60% (G +C), and careshould be taken to
denatured. Initial denaturation of longer than 2 minutes at 95°C is unnecessary
avoid sequences that might produce inernal secondary structure. The 3 -ends of the
and may reduce yield.
primers should not be complementary to avoid the production of primer-dimers
C.The externsion time should be at least 1min/kb target length. Primer-dimers unnecessarily deplete primers from the reaction and result in an unwanted
Table 1. Recommended Thermal Cycling Conditions for GoTaq DNA polymerase reaction that competes with the desired reaction. Avoid three G or C
Polymerase-Mediated PCR Amplification. These guidelines are optimal nucleotides in a row near the 3-end of the primer, as this may result in nonspecific
for the Perkin Elmer thermal cycler model 480 or comparable thermal cyclers. primer annealing, increasing the synthesis of undesirable reaction products. Ideally, both
primers should have nearly identical melting temperatures (T): in this manner, the two
Step Temperature Time Number of Cycles
primers should anneal at roughly the same temperature. The annealing temperature of the
Initial Denaturation 95°C 2 minutes 1 cycle reaction depends on the primer with the lowest Tm For assistance with calculating the T
Denaturation 95°C 0.5-1 minute of any prime, a ImCalculator is provided on the BioMath page of the Promega wetb site

Annealing 42-65°C 0.5-1 minute 25-35 cycles at: www.promega.com/biomath/

Extension 72°C 1min/kb
D. Amplitication Troubleshooting
Final Extension 72°C 5 minutes cycle o overcome low yield or no yield in amplitications (eg., mouse tail genotyping
Soak 4°C Indefinite 1 cycle
applications), we recommend the following suggestions
"Annealing temperature should be optimized for each primer set based on the primer T
Adjust annealing temperature. The reaction buffer composition aftects the melting
tor
Separate the PCR products by agarose gel electrophoresis and visualize with ethidium properties of DNA. See BioMath Calculator to calculate the melting temperature
bromide or any other means. For reactions containing the 5X Green GoTaqo Reaction primers in the GoTaq reaction (www.promega.com/biomath/)
Buffer, load the amplification reaction onto the gel directly after amplification. Minimize the effect of amplification inhibitors. Some DNA isolation procedures,
Reactions containing the 5X Colorless Golaq Reaction Buffer also can be loaded particularly genomic DNA isolation, can result in copurification of amplification
will need to be added to
directly into the wells of an agarose gel, but a tracking dye inhibitors. Reduce the volume of template DNA in the reaction, or dilute template DNA
efective
monitor the progress of electrophoresis.
prior to addingto reaction. samples 1:10,000 has been shown to be
Diluting
In improving results, depending on initial DNA concentration.
2. General Considerations
and wash step prior to
Increase template DNA purity. Include an ethanol precipitation
A. Enzyme Concentration with the DNA.
amplification to remove inhibitors that copurity
We have found that 1.25 units of GoTag DNA Polymerase per 50ul amplification reaction DMSO or betaine) may
Add PCR additives. Adding PCR-enhancing agents (e.g,
is adequate for most amplifications. Adding extra enzyme generally does not produce such as BSA (Sigma Cat.f A7030, final
Improve yields. General stabilizing agents tailure.
significant increases in yield. However, in some cases,more enzyme may be beneficial. Concentration 0.16mg/ml) also may help to
overcome amplification
Please be aware that excessive amounts of enzyme and exoesSively long extension times
increase the likelihood of generating artifacts due to the intrinsic 5 3 exonuclease
activity of Taq DNA polymerase Part# 9PIM300
Printed in USA. Revised 4/18.

www.promega.com
Promega Corporation 2800 Woods Hollow Road-Madison, wi 53711-5399 U.S.A. Toll Free in the USA 800-356-9526
Telephone608-274-4330