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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FasTract®  Direct PCR Kit for Tissue

Guidelines of Sample Preparation

• When working with tissue samples, use of 2mg of tissue is recommended for direct PCR.
• Use a scalpel to cut out the tissue.
• The scalpel must be thoroughly cleaned with 2% Sodium hypochloride or 70% Ethanol to prevent cross contamination.
• Collect the tissue samples in a PCR tube, add 100µl Fastract Tissue Lysis Buffer into each tubes,  vortex well and incubate them at 95° 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 1.5µl of this clear lysate as template for the PCR reaction.

Guidelines for PCR Setup

• Add 47.5µl 1X Fastract Tissue cell Direct PCR mix in a Flat Capped PCR tube
• Add 0.5µl of each Primers (from 10mM stock) to the master mix
• Use1.5µl of Fastract cultured cell 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. 

Troubleshooting

• In case of no PCR reaction dilute the tissue lysate 1:10 or 1:50 with Fastract Tissue lysis buffer and use 1.5 μl of this dilution as template
• In case of non-specific amplification, increase the annealing temperature, reduce the total number of cycles or design a new set of primers.

Ordering Information

Product Details Specifications Protocols & Manual Product Resource

Description

FasTract® Tissue Direct PCR Kit is designed to perform PCR directly from non-fixed animal derived tissue samples tissue samples without prior DNA purification. Fresh  tissues or tissue samples frozen at -20°C are all suitable templates for this kit. The kit employs a specially engineered high fidelity DNA Polymerase enzyme that exhibits high resistance to many PCR inhibitors found in animal tissues. The kit ensures high yield and is time saving as DNA isolation from samples can be avoided prior to PCR. The kit includes a pair of universal control primers that is compatible with a number of vertebrate species .The kit is recommended for end-point PCR.

FasTract®  Direct PCR Kit for Tissue

• Fastract Tissue Lysis Buffer
• Fastract Tissue 1X 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 Tissue sample lysate last to the reaction.
• It is recommended to add the tissue samples into the empty tube and add the lysis buffer to ensure that the entire sample is dipped into buffer. Make sure that you see the sample in the solution.
• 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

Protocol & Manuals

Some product description and more information to download our product brochure or manual datasheet

Catalogue No.	Product Details	Datasheet	Brochure
G45314A	FasTract®  Direct PCR Kit for Tissue, 100 rxn
G45314	FasTract®  Direct PCR Kit for Tissue, 500 rxn