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Description

FasTract® Blood Direct PCR Kit is designed to perform PCR directly from blood, and saliva without prior DNA purification. Freshly collected blood, blood stored with anticoagulants, clotted blood and even blood spots on paper discs, all are suitable samples for this FasTract® Blood Direct PCR kit. The kit employs a specially engineered high fidelity DNA Polymerase enzyme that exhibits high resistance to many PCR inhibitors found in blood and other bio-fluids. The kit ensures high yield and is time saving as DNA isolation from samples can be avoided prior to PCR. The kit is recommended for end-point PCR.

FasTract® Direct PCR Kit for Blood is a rapid PCR mastermix that eliminates the requirement of genomic DNA isolation prior PCR reaction. This improved master mix is a ready load 2X mastermix, allowing researcher to load the PCR product directly in the gel. Addition of loading dye is not required. FasTract®  direct PCR kit comes with three different components.

1. RBC Lysis Buffer
2. Pellet dissolving buffer
3. 2X mastermix

This is recommended to lyse the RBC and harvest the WBC (protocol provided with the kit) which to be resuspended and lysed in pellet dissolving buffer. This lysate would be used as template for the PCR reaction. Fastract® amplification after removal of RBC is always recommended because it helps to have efficient amplification for long amplicons and also for multiplex amplification (Figure 1 and Figure 2). Direct lysis of whole blood followed by amplification with Fastract® PCR master mix shows efficient PCR reaction for short amplicons (less than 500bp) but the same for long amplicons (more than 1 kb) are compromised (Figure 3 and 4 respectively). Amplification efficiency from blood with nucleated RBC's are also checked and efficient amplification were observed (Figure 5). The pellet dissolving buffer has been formulated in such a way, lysate would be compatible to amplify with any market leading Taq DNA Polymerase (Figure 6).

FasTract® Direct PCR Kit for Blood

• FasTract® Blood Buffer
• FasTract® 2X PCR Mix (FasTract® DNA Polymerase, Reaction buffer, dNTP (10mM))
• Control Primer Mix (10mM each)

Important Notes

• Carefully mix and spin down all tubes before opening to ensure homogeneity and improve recovery. The PCR setup can be performed at room temperature.
• Always add the Blood lysate last to the reaction.
• When working with clotted blood vortex vigorously after addition of lysis buffer
• Always ensure that the volume of lysate that is being added to the PCR reaction is 3% of the total reaction volume. Do not add more lysate to the reaction mix

Shipping and Storage

FasTract® tissue Direct PCR Kit is shipped on gel ice. Upon arrival, store the components at  4°C. Do not freeze.

Protocol & Manuals

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