2X Hot Start XtraWhite Master Mix GL and 2X Hot Start XtraRTL Master Mix GL (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). 2X Master Mix Standard GL contain an internal fluorescent stain for DNA detection on Agarose gel directly after PCR amplification.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 GL 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 GL 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.
GREEN LINE OVERVIEW
2X Master Mix GL contains an internal fluorescent stain for DNA detection on Agarose gel directly after PCR amplification.
cat. no.
amount
note
cat. no.
amount
note
STS-HXMMixW 200 GL*
5 ml
2X Hot StartXtra White Master Mix GL
STS-HXMMixRTL 200 GL
5 ml
2X HotStart Xtra RTL Master Mix GL
STS-HXMMixW 1000 GL*
25 ml
*2X XtraWhite Master Mix is supplied with appropriate quantity of 6X Loading Dye
2X Hot StartXtra White Master Mix GL
STS-HXMMixRTL 1000 GL
25 ml
2X HotStart Xtra RTL Master Mix GL
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 HSMaster Mix GL
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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