Ruby Hot Start Master (2x)
Detailní popis
Red master mix for highly sensitive and specific PCR, direct gel loading
Ready-to-Use Mixes for PCR
For general laboratory use.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
Short term storage (up to 3 month) at 4 °C possible.
Shelf Life: 12 months
Form: liquid
Concentration: 2x conc.
Description:
Ruby Hot Start Master is a 2 x conc. ready-to-use master mix with blocked polymerase activity at ambient temperature.
The master mix is recommended for routine PCR applications, high throughput PCR or genotyping and provides an improved specificity and sensitivity when amplifying low-copy-number targets, working with complex backgrounds or when prolonged room-temperature set-ups are unavoidable.
The thermal activation at the onset of the initial denaturation prevents the extension of nonspecifically annealed primers and primer-dimer formation at low temperatures during PCR setup.
It contains all reagents required for PCR (except template and primer) in a well-balanced ratio to ensure high specificity and minimal by-product formation in almost all PCR applications without the need of additional optimization steps.
Ruby Hot Start Master contains an inherent red dye and gel loading buffer allowing an easy visual control during PCR set-up and the direct loading of the reaction product into the gel.
The mix guarantees robust and reliable amplification results with a minimum of pipetting steps, saves time and reduces the risk of contaminations.
The total PCR assay volume is freely adaptable to individual protocols or the requirements of automated pipetting systems.
Content:
| Cat.No. | Master Mix | PCR-grade water | Assays x 50 μl |
| PCR-165S | 4 x 1.25 ml | 6 ml | 200 |
| PCR-165L | 20 x 1.25 ml | 2 x 12 ml | 1000 |
| PCR-165XL | 100 ml | 100 ml | 4000 |
2 x concentrated PCR master mix containing aptamer inhibited hot start Taq polymerase, nucleotides (dATP, dCTP, dGTP, dTTP), KCl, (NH4)2SO4, MgCl2, red dye, density reagent, enhancing and stabilizing additives.
Recommended PCR assay:
Before starting, vortex the master mix thoroughly to assure homogeneity.
| component | stock. conc. | 20 μl assay |
50 μl assay |
final conc. |
| Ruby Hot Start Master | 2x | 10 μl | 25μl | 1x |
| Primer Mix or each primer |
10 μM each primer | 0.4-0.8 μl | 1-2 μl | 200-400 nM each primer |
| Template/ sample DNA | < 10 ng | < 20 ng | ||
| PCR-grade water | fill up to 20 μl | fill up to 50 μl |
Recommended cycling conditions:
Before cycling, vortex PCR tubes or plates to assure homogeneity and centrifuge briefly to remove bubbles.
| Initial denaturation |
95 °C | 2 min | 1x |
| Denaturation Annealing1) Elongation2) |
95 °C 50 - 68 °C 72 °C |
10 - 20 sec 10 - 20 sec 20 sec - 4 min |
25 - 30x |
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.
BIOZ Product Citations:
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