Extract-N-Amp Plant Flow Chart

Description

The Extract-N-Amp™ Plant PCR Kits contain all the reagents necessary to rapidly extract genomic DNA from plant leaves and amplify targets of interest by PCR.

  • Novel – single-step extraction of genomic DNA to PCR
  • Fast – tissue to PCR in 15 minutes
  • Convenient – no long enzymatic digestions

The Extract-N-Amp™ Plant PCR Kit offers a novel Extraction Solution that eliminates the need for freezing of plant tissues with liquid nitrogen, mechanical disruption, organic extraction, column DNA purification, or alcohol precipitation. The product includes a specially formulated hot start PCR ReadyMix for amplification directly from the extract. Extract-N-Amp™ products are Advanced Technology certified.

Procedure

Genomic DNA is extracted from 0.5 to 0.7 cm plant leaf disks that have been cut with a standard paper punch and simply incubated in Extraction Solution at 95 °C for 10 minutes. An equal volume of Dilution Solution is added to the extract to neutralize inhibitory substances prior to PCR. A portion of the DNA extract is then added directly to the optimized PCR mix supplied.

Application

Perfect for genotyping.

Features and Benefits

Pick the formulation that is right for you

The Extract-N-Amp™ Tissue PCR Kits include a specially formulated hot start PCR ReadyMix for amplification directly from the extract.

The PCR ReadyMix comes in two formulations:

  • REDExtract-N-Amp™ PCR Ready Mix – contains an inert dye that acts as a tracking dye and allows for convenient loading of PCR reactions onto agarose gels for analysis.
  • Extract-N-Amp™ PCR Ready Mix – no dye contained.

Order Information

Extract-N-Amp™ Plant PCR Kits
REDExtract-N-Amp™ Plant PCR Kits

Kit Contents

Extract-N-Amp™ Kit

  • Extraction Solution
  • Dilution Solution
  • Extract-N-Amp™ PCR Ready Mix
  • 2 mL Tubes for Extraction (not included with the 1,000 Amplification kit)

REDExtract-N-Amp™ Kit

  • Extraction Solution
  • Dilution Solution
  • REDExtract-N-Amp™ PCR Ready Mix
  • 2 mL Tubes for Extraction

Sample Data

PCR analysis of genomic DNA extracted from 5 different plant species using our Extract-N-Amp™ Plant Kit

Figure 1. Genomic DNA was extracted from 0.5 cm leaf disks that were cut using a standard paper punch. DNA was extracted using the Extract-N-Amp™ Plant PCR Kit in less than 15 minutes. All samples were then amplified using the specially formulated Hot Start PCR mix. The products were generated from a 30-cycle duplex reaction containing primers specific to plant chloroplast (upper band) and primers specific to Cannabis sativa DNA (lower band). MW ladder is 100, 200, 400 and 800 bp. Data provided by Andy Hopwood, Forensic Science Service, Birmingham, England.

Sequence was resolved on an ABI 310 from a purified, 645 bp corn leaf PCR product

Click on image to enlarge.

Figure 2. The PCR product was purified with the GenElute™ PCR Clean-Up Kit (Product No. NA1020). The DNA extraction and PCR were performed using our Extract-N-Amp™ Plant PCR Kit. The sequence was obtained using the same primers as for the original PCR.

Stability of Extract-N-Amp™ Plant Extracts

Figure 3. Eight disks were punched from a corn leaf, and DNA was extracted according to the procedure in the Technical Bulletin for the Extract-N-Amp™ Plant Kit. Two 4 µL aliquots from each were analyzed immediately by quantitative PCR with SYBR® Green detection on an ABI Prism 7700. DNA standards for quantitative PCR were purified DNA prepared from corn leaf tissue with the GenElute™ Plant Genomic DNA Kit (Product No. G2N70). Half of the leaf extracts were stored at 4 °C (recommended storage conditions) and the other half at 37 °C (accelerated storage). Quantitative PCR was repeated after 1 week, 3 weeks, 6 weeks and 6 months from extracts at 37 °C. Results for storage at 37 °C are shown. The average of 2 replicate PCR assays from each extract is plotted. Error bars represent one standard deviation.

extract-n-amp-chilli

PCR results from Extract-N-Amp™ plant preparation of chilli 0.5cm diameter leaf punch
4 µL of the elution was used for 20 µL PCR reaction mix. Capsicum frutescens fasciculate (FA) gene Primers used. Lane (1) D7058 DirectLoad™ Wide Range DNA Marker; (2) Chilli Leaf DNA from Extract-N-Amp™, Sample 1; (3) Chilli Leaf DNA from Extract-N-Amp™, Sample 2; (4) No template control; (5) Chilli DNA from CTAB Extraction.

xna-chilli-seed

PCR results from Extract-N-Amp™ Seed preparation of Chilli seeds
4 µL of the elution was used for 20 µL PCR reaction mix. Capsicum frutescens fasciculate (FA) gene Primers used. Lane (1) D7058 DirectLoad™ Wide Range DNA Marker; (2) Chilli seed DNA from Extract-N-Amp™, Sample 1; (3) Chilli seed DNA from Extract-N-Amp™, Sample 2; (4) No template control; (5) Chilli seed DNA from CTAB Extraction.

xna-eggplant-seed

PCR results from Extract-N-Amp™ Seed preparation of Brinjal (eggplant) seeds
4 µL of the elution was used for 20 µL PCR reaction mix. Solanum melalogena Cytochrome P-450EG4 Primers used. Lane (1) D7058 DirectLoad™ Wide Range DNA Marker; (2) Brinjal seed DNA from Extract-N-Amp™, Sample 1; (3) Brinjal seed DNA from Extract-N-Amp™, Sample 2; (4) No template control; (5) Brinjal seed DNA from CTAB Extraction.

xna-tobacco-seed

PCR results from Extract-N-Amp™ Seed preparation of Tobacco seeds
4 µL of the elution was used for 20 µL PCR reaction mix. Nicotiana tabacum RbcS transit peptide primers used. Lane (1) D7058 DirectLoad™ Wide Range DNA Marker; (2) Tobacco seed DNA from Extract-N-Amp™, Sample 1; (3) Tobacco seed DNA from Extract-N-Amp™, Sample 2; (4) No template control; (5) Tobacco seed DNA from CTAB Extraction.

xna-rice-seed

PCR results from Extract-N-Amp™ Seed preparation of Rice seeds
4 µL of the elution was used for 20 µL PCR reaction mix. Rice actin primers used. Lane (1) 1 Kb DNA Ladder; (2) Rice seed DNA from Extract-N-Amp, Sample 1; (3) Rice seed DNA from Extract-N-Amp, Sample 2; (4) No template control; (5) Rice seed DNA from CTAB Extraction.

xna-tobacco

PCR results from Extract-N-Amp™ plant preparation of Tobacco 0.5cm diameter leaf punch
4 µL of the elution was used for 20 µL PCR reaction mix. Nicotiana tabacum RbcS transit peptide gene Primers used. Lane (1) D7058 DirectLoad™ Wide Range DNA Marker; (2) Tobacco Leaf DNA from Extract-N-Amp™, Sample 1; (3) Tobacco Leaf DNA from Extract-N-Amp™, Sample 2; (4) No template control; (5) Tobacco DNA from CTAB Extraction.

xna-cotton-seed

PCR results from Extract-N-Amp™ Seed preparation of Cotton seed embryo
4 µL of the elution was used for 20 µL PCR reaction mix. Cotton rbcs promoter gene primers used. Lane (1) 1 Kb DNA Ladder; (2) Cotton seed DNA from Extract-N-Amp™, Sample 1; (3) Cotton seed DNA from Extract-N-Amp™, Sample 2; (4) No template control; (5) Cotton seed DNA from CTAB Extraction.

Materials
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