Fastract® Direct PCR Kit for Blood
Fastract Direct PCR Kit®
The kit employs a specially engineered high fidelity DNA
Polymerase enzyme that exhibits high resistance to many PCR inhibitors
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Find support for technicalities of applications and product use
Catalogue No. : G4667
Pack Size: 50 Preps
FasTract® Direct PCR Kit for Blood
Guidelines of Sample Preparation
- Blood stored with anticoagulants, clotted blood and even blood spots on paper discs, all are suitable samples for this Fastract® Blood Direct PCR kit.
- When working with clotted blood or blood, 10-20 µl blood is recommended.
- In case of spotted blood on paper discs use a single hole paper puncher to punch out a spotted blood disc. This will be the sample.
- Collect the blood samples in a PCR tube , add 100µl Fastract® Blood Lysis Buffer into each tubes, vortex well and incubate them at 95°C for 15 mins.
- After incubation centrifuge the PCR tubes at 15000 rpm for 2 mins. Collect the clear supernatant in a separate microfuge tube.
- Use 1.5 µl of this clear lysate as template for the PCR reaction.
Guidelines of PCR setup
- Add 47.5 µl1X FasTract® Blood Direct PCR mix in a Flat Capped PCR tube.
- Add 0.5 µl of each Primers (from 10mM stock) to the master mix.
- Use1. 5 µl of FasTract® Blood lysate as template for the PCR reaction.
- While setting up the PCR cycle keep the initial denaturation time to be 5 mins at 95°C followed by denaturation at 95°C for 10 -15 sec and annealing at the desired Tm. For extension, use 1min for 1Kb amplicon size.
In case of PCR reaction dilute the lysate and case of non-specific amplification, increase the annealing temperature, reduce the total number of cycles or design a new set of primers.
|Cat. No.||Product||Pack Size||Price|
|G45312A||FasTract® Direct PCR Kit for Blood||100 rxn||₹ 10,250|
|G45312B||FasTract® Direct PCR Kit for Blood||500 rxn||₹ 28,360|
|G45312||FasTract® Direct PCR Kit for Blood||1000 rxn||₹ 51,000|
|G45312Q||FasTract® Blood lysis Buffer for Blood (Without Mastermix)||1000 rxn||₹ 44,210|
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.
- RBC Lysis Buffer
- Pellet dissolving buffer
- 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)
- 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
Some product description and more information to download our product brochure or manual datasheet
|Catalogue No.||Product Details||Datasheet||Brochure|
|G45312A||FasTract® Direct PCR Kit for Blood,100 rxn|
|G45312B||FasTract® Direct PCR Kit for Blood,500 rxn|
|G45312||FasTract® Direct PCR Kit for Blood,1000 rxn|
|G45312Q||FasTract® Blood lysis Buffer for Blood (Without Mastermix), 1000 rxn|