Catalog No. : MN-740770.50
NucleoSpin Plant II (50)
|NucleoSpin Plant II (50)||$243.00|
The second generation for rapid isolation of genomic DNA from plant samples
with higher yield and quality
• Two alternative lysis buffers included for optimal processing of various samples
and highest flexibility:
Lysis Buffer PL1, based on CTAB lysis method
Lysis Buffer PL2, based on SDS lysis method
• NucleoSpin® Filters included for clarification of lysate
• Improved buffer compositions – higher yield and purity
• Optimized silica membrane – improved DNA binding
• RNase A included
|Format||Mini spin columns|
< 100 mg plant tissue (wet weight), < 20 mg plant tissue (dry weight)
|Lysate clarification||NucleoSpin® Filters|
|Fragment size||50 bp–approx. 50 kbp|
|Typical yield||1–30 µg (100 mg plant tissue, wet weight)|
|Elution volume||50–100 µL|
|Preparation time||30 min/prep|
|Binding capacity||50 µg|
Principle / Procedure
With NucleoSpin® Plant II MACHEREY-NAGEL introduces the second generation of kits for isolation of gDNA from plant material, fungi, and other biological samples.
The kit includes two optimized, alternative lysis buffers based on the established CTAB and SDS lysis methods. As plants are very heterogenous and contain a lot of different metabolites like polyphenols, polysaccharides, or acidic components, NucleoSpin® Plant II offers two different lysis procedures for optimal processing of various samples.
The silica membrane in the NucleoSpin® Plant II Columns is optimized to improve DNA binding. NucleoSpin® Filters are included for convenient clarification of lysates.
NucleoSpin® Plant II allows processing of up to 100 mg (wet weight) or 20 mg (dry weight) starting material. Depending on the individual sample typical yields are in the range from 1 to 30 μg DNA. The eluted DNA is ready to use for subsequent reactions like PCR, restriction analysis, and others.
PCR-grade DNA from different Orchidaceae species
Genomic DNA of four different Orchidaceae species was isolated with NucleoSpin® Plant II using Lysis Buffer PL1 (procedure performed without the optional wash step). The starting material was 20 mg silica-gel dried plant tissue, the homogenization of the plant material was performed with a bead mill (Retsch).
Isolated DNA was subjected to PCR (DNA concentration: 10 ng/μL) with two chloroplast-specific primer systems and analyzed on a 1 % TAE agarose gel.
M = marker (l-HindIII)
A: PCR reaction probing for matK (1800 bp fragment)
B: PCR reaction probing for trnL-trnF (1200 bp fragment)
NucleoSpin® Plant II allows purification of amplifiable DNA from dried plant samples. All samples show the expected signal, PCR inhibition was not observed.
Data kindly provided by Dr. A. Kocyan, Systematic Botany, Ludwig Maximilian University Munich, Germany
1. Stereochilus sp. (leaf)
2. Doritis pulcherrima (leaf)
3. Cleisostoma racemiferum (inflorescence rachis)
4. Trichoglottis sp. (leaf)
Comparison to competitor kits
100 mg of fresh fir needles (Abies alba) have been processed using the NucleoSpin® Plant II kit (Lysis Buffer PL1 and Lysis Buffer PL2 tested) and competitor products (Q, P, and I). 10 μL of DNA eluate were analyzed on a 1 % TAE agarose gel.
Fir needles are known to contain large amounts of hydrophobic compounds that may negatively influence purity of DNA.
For this sample material highest yields and best purity of DNA could be obtained with the NucleoSpin® Plant II lysis buffer PL2.
Note: The experimental results show that with this sample material there is a significant difference between the performance of the two lysis buffers included in the NucleoSpin® Plant II kit. Testing of both lysis buffers is therefore recommended.
PCR-grade DNA from herbarium samples
Genomic DNA of five different Brassicaceae species was isolated with NucleoSpin® Plant II using Lysis Buffers PL1 (A) and Lysis Buffer PL2 (B) and with a kit of competitor I (C). The starting material was 8–20 mg dried plant leaf (herbarium material), homogenization was performed with a mortar and pestle in the presence of sea sand and the respective lysis buffer.
Isolated DNA was subjected to PCR (2 μL out of 50 μL eluate used in a 50 μL PCR reaction) with an ITS-specific primer system (ITS, internal transcribed spacer, part of the ribosomal DNA). The 740 bp PCR product was analyzed on a 1.5 % TBE agarose gel (M = marker HyperLadder II, Bioline).
1. Isatis kotschyana
2. Sameraria nummularia (difficult to process)
3. Myagrum perfoliatum
4. Tauscheria lasiocarpum
5. Boreava orientalis
NucleoSpin® Plant II Lysis Buffers PL1 and Lysis Buffer PL2 allow purification of amplifiable DNA even from difficult-to-process, dried plant samples. All of the samples show the expected signal, PCR inhibition was not observed whereas competitor I fails to provide PCR-grade DNA from sample 2.
Data kindly provided by U. Coja, Systematic Botany, University of Osnabrück, Germany
Proven suitability of lysis buffers – plant species tested with NucleoSpin® Plant II
This table gives an overview about customer data on different plant species that have been tested using NucleoSpin Plant II. It indicates plant species that were successfully tested and the corresponding buffer system that was used (ü).
Important! For a large variety of plant species both lysis buffers will give good results. Use the table only for rough orientation and guideline which buffer system has already been tested. In order to obtain optimal results with your individual sample material we recommend to test both buffers in parallel to check which system will be suited best!