Fastract® Direct PCR Kit

The kit employs a specially engineered high fidelity DNA
Polymerase enzyme that exhibits high resistance to many PCR inhibitors

• GSure® Ultra Nucleic Acid Kit for DNA & RNA Isolation
• GSure® Dogma kit for DNA, RNA & Protein Isolation
• Reverse Transcription & cDNA Synthesis
• Thermostable DNA Polymerases
• Fastract® Direct PCR Kit
• SYBR green & Taqman based qRT-PCR Kits
• TA-Cloning kit and Related Enzymes
• Competent Cells
• Site Directed Mutagenesis Based Kits
• equencing Grade Reagents

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  Fastract® Direct PCR Kit for Plant

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Guidelines of Sample Preparation

• Single hole paper puncher must be used to obtain small and uniform leaf discs (2mm diameter)
• It is very important to clean the cutting edge every time before sampling to prevent cross contamination between samples
• 2 % sodium hypochlorite solution or 70% Ethanol should be used for cleaning.
• Punch out a disc from the plant leaf using the sampling tool and place disc directly into the PCR tubes.
• Add 50µl FasTract® Lysis Buffer into each tubes, vortex well and incubate them at 95°C for 15 mins.
• Use 2. 5µl of this lysate as template for the PCR reaction
• The FasTract® Lysis buffer can be used for collection and storage of leaf disc samples.

 

Guidelines for Control reactions

We recommend setting up a control PCR reactions with both the purified DNA (provided) as well as the Fastract® leaf disc lysate used in the actual experiment to ensure that the PCR conditions are optimal. If the positive control with purified DNA fails, the PCR conditions should be optimized before continuing further.It is recommended to add a no-template control to all PCR assays.

 

Guidelines For PCR setup

• Take a single leaf disc in a PCR tube and add 50µl of Fastract® Lysis Buffer in it.
• For samples collected directly in Fastract® Lysis buffer, next step is not required for soft plant leaves. Still, heating the samples will ensure better lysis.
• Vortex the sample vigorously and incubate the  Fastract® Lysis Buffer added sample at 95°C for 15 min in a PCR machine lead heating condition.
• Add 22µl 1X Fastract® PCR mix in a Flat capped PCR tube.
• Add 0.25µl of each Primers (from 10mM stock) to the master mix.
• Use 2. 5µl of Fastract® lysate as template for the PCR reaction.
• Set up PCR as below.

 

Troubleshooting

• For tough and dry leaf samples increase the o incubation time at 95°C.
• If PCR fails due to high amount of PCR inhibitors in the leaf disc lysate, then the amount of lysis buffer can be increased to 75-100µl.
• If PCR fails even after increasing the volume of lysis buffer, incubate half of the leaf disc in lysis buffer instead of the entire disc.

 

Ordering Information

Cat. No.	Product	Pack Size	Price
G45311A	Fastract® Direct PCR Kit for Plant	100 rxn	POR
G45311B	Fastract® Direct PCR Kit for Plant	500 rxn	POR

 

Product Details Specifications Protocols & Manual Product Resource

 

Introduction

Plant DNA isolation is a time consuming, laborious procedure specially when working with a huge number of samples. Fastract® Plant Direct PCR Kit is designed to perform PCR directly from plant leaves without prior DNA isolation and purification. Fresh plants, plant material stored at 4°C or frozen are all suitable templates for this kit. Plant leaves could also be collected directly in the Fastract® plant Lysis Buffer and stored at room temperature for at least 48hr, at 4°C for 5 days and at -20°C for at least one month. Chaotropes and detergents present in Fastract® Lysis Buffer ensure better and efficient lysis even with though samples like palm and coconut. Fastract® plant PCR Mix contains a genetically modified Hi-Fidelity DNA polymerase which can amplify even in presence of PCR inhibitors in plant lysate. The kit ensures high yield in PCR amplification and is time saving as DNA isolation from samples can be avoided prior to PCR. The control primers provided can amplify a highly conserved region from plant DNA. Purified plant gDNA is also provided as a positive control for the PCR reactions. The kit is recommended for end-point PCR.

 

Fastract® Direct PCR Kit for Plant

 

 

 

 

 

 

• Fastract® Plant 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 plant sample last to the reaction.
• It is recommended to eject the leaf disc into the empty tube and add the lysis buffer to ensure that the entire disc is dipped into buffer. Make sure that you see the sample disc in the solution.
• We recommend using fresh plant material for best results, even though plant material stored at  4°C or -20°C can also be used.
• For extension, use 1min for 1Kb amplicon size.
• The Fastract® Lysate must be added such that it is 10% of the total reaction volume i.e if the total reaction volume is 25 µl then 2.5 µl of leaf disc lysate must be added to the reaction.

Shipping and Storage

Fastract® 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
G45311A	Fastract® Direct PCR Kit for Plant, 100 rxn
G45311B	Fastract® Direct PCR Kit for Plant, 500 rxn