2X Hot Start XtraWhite Master Mix and 2X Hot Start XtraRTL Master Mix (GeneSpin proprietary formulation), are two premixed, ready-to-use solution containing Hot Taq Pol, dNTPs and MgCl2 in a Reaction Buffer optimized for use in PCR amplification of targets present in low copy number and to avoid amplification of non-specific products. Both buffers contain 3.0mM magnesium, PCR enhancers and thickening agents (vortex thoroughly prior to use).
Both Master Mixes allow for specific PCR amplification by keeping the enzyme inactive until the temperature reaches approximately 40°C, while also reducing samples preparation time as well as risk of contamination from multiple pipetting steps.
2X Hot Start XtraRTL Master Mix contains Orange G dye that allows gel loading and electrophoresis of the sample directly from the PCR* tube, without further manipulation. 2X Hot Start XtraWhite Master Mix is supplied with appropriate quantity of 6X Loading Dye. The Orange G dye migrates at the same rate as a duplex DNA fragment of approximately 50 Kbp and does not interfere with DNA migration when it is used as a loading dye for agarose gel electrophoresis.
cat. no.
amount
note
cat. no.
amount
note
5 ml
25 ml
*2X HotStart Xtra White Master Mixis supplied with appropriate quantity of 6X Loading Dye
2X HotStart Xtra White Master Mix
2X HotStart Xtra White Master Mix
5 ml
25 ml
2X HotStart Xtra RTL Master Mix
2X HotStart Xtra RTL Master Mix
FOR RESEARCH USE ONLY
SHIPPING
Shipped in green ice.
FORM
Liquid
SHELF LIFE
12 months
UNIT DEFINITION
One unit is defined as the amount of enzyme required to incorporate 10 nanomoles of dNTPs into acid-insolubile material in 30 min at 74°C.
CONCENTRATION
2X conc.
STORAGE
Store at -20°C
Assay set up
Before starting, vortex all components thoroughly to ensure homogeneity.
Prepare a premix for the number of assays you need according to the following protocol:
component
stock conc.
final conc.
30ul reaction
2X Master Mix
primers
DNA Template
2X
-
1ug/ul each
1X
50ng/ul each
-
3.0ul
2.0ul each
10-20ng
MG Water
-
-
up to 30ul
Cycling conditions
Cycling conditions:
Spin down the tubes/plate briefly to remove bubbles and place them into the cycler.
denaturation
95°C
5min
denaturation (1)
95°C
30 sec
annealing (2)
extension
50-68°C
72°C
30 sec
30 sec
final elong.
72°C
5 min
final step
4°C
forever
1)The annealing temperature depends on the melting temperature of the primers used.
2)The elongation time depends on the length of the fragments to be amplified. A time of 1 min/kb is recommended.
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yours
MOLECULAR BIOLOGY
reagents
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