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Hot Start Master Lyophilisate

Lyophilized Hot Start Master Mix

Product Cat. No. Amount Price (EUR) Buy / Note
preloaded 8-tube strips (high profile) PCR-153S-8TS 12 strips (96 reactions) 120,00 Add to Basket/Quote Add to Notepad
preloaded 8-tube strips (high profile) PCR-153L-8TS 60 strips (480 reactions) 480,00 Add to Basket/Quote Add to Notepad
preloaded 96-well plates (flat top / without skirt) / high profile PCR-153S-FTP 2 plates (192 reactions) 180,00 Add to Basket/Quote Add to Notepad
preloaded 96-well plates (flat top / without skirt) / high profile PCR-153L-FTP 10 plates (960 reactions) 720,00 Add to Basket/Quote Add to Notepad
preloaded 96-well plates (half skirt) / high profile PCR-153S-HSP 2 plates (192 reactions) 180,00 Add to Basket/Quote Add to Notepad
preloaded 96-well plates (half skirt) / high profile PCR-153L-HSP 10 plates (960 reactions) 720,00 Add to Basket/Quote Add to Notepad

For in vitro use only!

Shipping: shipped at ambient temperature

Storage Conditions: store at ambient temperature
Store in an aluminium-coated bag or on a dry place.
Lyophilisates may hydrate at humidity levels >70 % when sealing is opened.

Shelf Life: 12 months

Description:
Hot Start Master Lyophilisate is delivered in PCR reaction tube strips or 96-well plates preloaded with a complete hot start master mix in a dry, room temperature stable format. The lyophilisate combines highest performance with convenience of use and stability. There is no need for freezing, thawing or pipetting on ice. The few remaining pipetting steps minimize the risk of errors or contaminations.
Each vial contains polymerase, dNTPs and reaction buffer with MgCl2 required for a 20 μl PCR assay.
To perform PCR, fill up the vials with a premix of primers and PCR-grade water and add DNA template. If necessary, centrifuge to remove bubbles, vortex the vials to assure homogeneity and start cycling.
The Hot Start Master Lyophilisate provides improved specificity and sensitivity when amplifying low-copy-number targets in complex backgrounds. The polymerase activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of nonspecifically annealed primers and primer-dimers formed during PCR setup.

Kit contents:
Hot Start Master Lyophilisate
Preloaded lyophilisates of hot start DNA polymerase, dATP, dCTP, dGTP, dTTP, Reaction Buffer with MgCl2 and stabilizers.

PCR grade water

Activation step
Hot Start Master Lyophilisate requires no prolonged heating or denaturing step. The polymerase inhibiting ligand is quickly released at the increased temperature of thermal cycling.

Recommended 20 μl PCR assay:

forward Primer0.2 - 1 μM
reverse Primer0.2 - 1 μM
template DNA1 - 50 ng
PCR-grade waterfill up to 20 μl


Recommended cycling conditions:

Initial
denaturation
94 °C2 min1x
Denaturation94 °C30 sec30x
Annealing1)45 - 68 °C30 sec30x
Elongation2)72 °C30 sec - 3 min30x
Final
elongation
72 °C2 min1x
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.

For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.