DirectBlood Genotyping PCR Kit (500reactions)

Product#: 5000M
$1,438.40
Availability:
Ships in 1-2 weeks

DirectBlood Genotyping PCR Kit (500reactions)

Cat. No. 5000M
Size 1 kit
Store 

The freeze-dried DirectBlood Genotyping PCR Lyocake can be stored at room-temperature. Please store resuspended master mixes (2x) and primer/probe mixes (4x) at -20°C. Minimize the number of freeze-thaw cycles by storing in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4°C. Please store the included rehydration buffer and additional primer/probe mixes upon arrival at -20°C.

Description

DirectBlood Genotyping PCR Kit is a novelty on the market. It allows genotyping in real-time and without any previous DNA isolation. You simply save and money: directly use EDTA blood without any extraction steps.

The DirectBlood Genotyping PCR Kit is optimized to function with a wide range of different SNPs to detect possible nucleotide variations. It has been successfully tested with a variety of hydrolysis probes and hybridization probes.

The setup of a genotyping experiment can not be simpler anymore: Mix primers&probes with the Genotying mastermix, add the respective blood sample and start real-time PCR, this is it. 

It is even more simple: store the DirectBlood Genotyping PCR Kit at room-temperature. A cool chain is not needed anymore. The included master mix is freeze-dried and dissolves within a few seconds after addition of the rehydration buffer.

Additionally, we have already established SNP-assays for you. In case, you can not find your respective SNPs, please provide us with the rs-number of the SNP-assays your are interested-in and you will receive an individual quotation from us.

Content

DirectBlood Genotyping PCR Kit contains all components necessary for direct PCR or PCR based genotyping directly from blood samples including an engineered DNA polymerase, an specifically optimized reaction buffer system, and ultrapure dNTPs. An aptamer-based hot-start formulation of the included blood-resistant DNA polymerase prevents false amplification. Temperatures above 50°-55°C cause the aptamer’s secondary structure to melt and will set-free the polymerase. Only target specific primers and probes need to be added.

Quality Control Assays
DirectBlood Genotyping PCR Kit is tested for functional and accurate genotyping PCRs direct from EDTA blood samples. DNA polymerase activity: DNA polymerase activity has been monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and a DNA primer. Enzyme-concentration has been determined by proteinspecific staining. Please inquire more information at info@mypols.de for the lot-specific concentration. No contamination has been detected in standard test reactions.
Safety
This product does not require a Material Safety Data Sheet because it does neither contain more than 1% of a component classified as dangerous or hazardous nor more than 0.1% of a component classified as carcinogenic. However, we generally recommend the use of gloves, lab coats and eye protection when working with these or any other chemical reagents. myPOLS Biotec takes no liability for damage resulting from handling or contact with this product. Further information can be found in the REGULATION (EC) No 1272/2008 OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL.

Real-time PCR detection setup
The DirectBlood Genotyping Kit is compatible with a wide range of assays. It has been successfully tested with a variety of hydrolysis probes and hybridisation probes.

Preparations before use
Please rehydrate the lyocake by adding exactly 218µl of the respective Rehydration Buffer to the PCR mix, resulting in 250µl of ready-to-use 2x Master Mix. Subsequently invert the closed tube a few times or briefly vortex and spin down the mixture before use. Tube should be placed on ice after rehydration. The rehydrated 2x DirectBlood Genotyping PCR Mix is now ready to be used for setting up a PCR experiment or can be stored at -20°C.

Sample preparation
Dilute a given EDTA blood sample to 2% (v/v) in DNase-free water by adding of 10 µL blood to 490 µL water. Subsequently invert the closed tube a few times or briefly vortex the mixture before use. Samples can also be stored for a couple of weeks at 4°C, e.g., for re-testing purposes.

Cycler compatibility
This product is compatible for the use with any qPCR cycler not requiring a passive reference dye (not included in the kit).

Recommendations for sample handling
  • Spin down and mix all solutions carefully before use.
  • Include a control without sample as a negative control.
  • Minimize the number of freeze-thaw cycles by storing the rehydrated product in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4°C.
  • We recommend the use of disposable gloves, DNase and RNase free filter tips and plastics.
PCR Reaction Setup
 

Component

Volume

Final concentration

2x DirectBlood Genotyping PCR Mix

10 µL

1x

4x Primer/Probe Mix

5 µL

1x

Diluted blood sample (2%)

5 µL

0.5%

Total volume/reaction

20 µL

 


Suggested PCR protocol

 

 

 

Initial denaturation

95°C

2 min

Denaturation

95°C

10 sec

Annealing/Extension*

60°C

40 sec          (50 cycles)

* Fluorogenic Data should be collected during this step. Real-time PCR signals may be reduced to 25% of the signals obtained from extracted DNA samples. This is a normal result and will still be leading to significant, valid Ct-values and unambiguous melting peaks.

Please note
Suggested cycling protocols can vary and depend strongly on the cycler and applied PCR assay.

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