You are on page 1of 1

For research purpose only.

Not for use in diagnostic procedures for


clinical purposes. For IN VITRO USE ONLY.

i-Master mix PCR Kit


Cat. No. 25201 96 Tubes
25202 500 Tubes
25203 5ml

DESCRIPTION
PROTOCOL
i-Master mix PCR Kit contains two tube system. Taq PCR tube of i-Master mix
PCR Kit contains Taq DNA polymerase and chemical stabilizer. The other 1. Add template DNA and primers into i-Master PCR tube.
tube contains dNTP, buffer and chemical stabilizer. Two tube system Note : Recommended volume of template and primer : 3㎕~5㎕
improves the stability and activity of i-TaqTM DNA polymerase. Appropriate amounts of template samples.
•cDNA : 0.5-10% of first RT reaction volume
STORAGE •Plasmid DNA : 10pg-100ng
Store at -20℃. i-Master mix solution should be stored at 4oC after receiving, •Genomic DNA : 0.2-2ug for single copy
and then stable for 6 months. If more then 6 months, must be eliquoted, and Note : Appropriate amounts of primers
then stored at –20℃. •Primer : 10-30uM each (sense and anti -sense)
2. Add i-Master mix Solution into the i-Master PCR tubes to a total volume
CHARACTERISTICS
of 20㎕.
-Ready to use with only template DNA and primers
Note : if you use 3? l of template DNA and primers, add 17㎕ i-Master
-Stable for over 1 year at -20 ℃ mix solution.
-Saving money and time
-Constructed as a reaction volume of 20㎕ 3. Dissolve the blue pellet by pipetting.
Note : If the mixture lets stand at RT for 1-2min after adding i-Master
KIT CONTENT mix solution, the pellet is easily dissolved.
•i-Master PCR tube 96 (500) tubes 4. (Option) Add mineral oil.
i-Master PCR tube (component)* Note : This step is unnecessary when using a thermal cycler that
employs a top heating method(general method).
i-Taq DNA Polymerase 2.5U
5. Perform PCR of samples.
Chemical stabilizer I 1X
6. Load samples on agarose gel without adding a loading-dye buffer and
Loading buffer 1X perform electrophoresis.
*Lyophilized under iNtRON’s instruction
SUGGESTED CYCLING PARAMETERS
•i-Master mix Solution 2 (9)ml

i-Master mix solution (component) PCR product size


PCR cycle Temp.
dNTPs 250uM each 100-500bp 500-1000bp 1Kb-5Kb
Tris-HCl(pH8.3) 10mM
Initial denaturation 94℃ 2min 2min 2min
KCl 50mM
Denaturation 94℃ 20sec 20sec 20sec
MgCl2 1.5mM
30-40 Annealing 50-65℃ 10sec 10sec 20sec
Chemical stabilizer II 1X Cycles
Extension 65-72℃ 20-30sec 40-50sec 1min/Kb
EXPERIMENTAL INFORMATION
M 1 2 3 4 5 6 7 8 9 10 11 Final extension 72 ℃ Optional. Normally, 2-5min

Note : The PCR process is covered by patents issued and applicable in certain
A countries. iNtRON Biotechnology does not encourage or support the unauthorized or
unlicensed use of the PCR process. Use of this product is recommended for persons
that either have a license to perform PCR or are not required to obtain a license.
B

Fig. 1. Comparision of heat-stability of i-TaqTM and i-Master mix PCR Kit


PCR performed on cloned gene with i-TaqTM and i-Master mix PCR Kit after
heating at 95 ℃ with various denaturation time.
< Panel A > i-TaqTM DNA Polymerase <Panel B > i-Master mix PCR Kit
Lane M, 1Kb DNA Ladder; lane 1, 20 min; lane 2, 30 min; lane 3, 40 min;
lane 4, 50 min; lane 5, 60 min; lane 6, 70 min; lane 7, 80 min; lane 8, 90
min; lane 9, 100 min; lane 10, 110 min; lane 11, 120 min

iNtRON BIOTECHNOLOGY
www.intronbio.com
info@intronbio.com