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Biomiga Soil gDNA Kit - GD2412-01

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Description :

Introduction

The EZgeneTM Soil gDNA Kit is designed for a rapid and reliable purification of high-quality genomic DNA from various soil samples. Up to 1 gram of soil samples can be processed in less than 1 hour. The system combines the reversible nucleic acid-binding properties of ezBindTM matrix with a propriety buffer system to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and hybridization techniques. There are no organic extractions thus reducing plastic waste and hands- on time to allow multiple samples to be processed in parallel.

In this procedure, soil sample is homogenized and then treated in a specially formulated buffer that contains detergent. Humic acid, proteins, polysaccharides, and other contaminants are subsequently precipitated after a heat-frozen step. DNA is further purified with a DNA spin-column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in water or low ionic strength Elution Buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Storage and Stability

All components of the EZgeneTM Soil gDNA Kit should be stored at 22oC-25oC. Under these conditions, DNA has successfully been purified and used for PCR after 12 months of storage. During shipment, or storage in cool ambient conditions, precipitates may form in some buffers. It is possible to dissolve such deposits by incubation the solution at 65oC.

Kit Contents

Catalog #

GD2412-00

GD2412-01

GD2412-02

Preps

4

50

250

ezBind DNA columns

4

50

250

2 mL collection tubes

8

100

500

Glass beads

1 g

12g

55g

DH reagent

1.2 mL

12 mL

60 mL

Buffer LX

5 mL

40 mL

200 mL

Buffer P2

1.5 mL

15 mL

60 mL

Buffer BL

3 mL

30 mL

150 mL

Elution Buffer

1.5 mL

20 mL

100 mL

DNA Wash Buffer

2 mL

15 mL

3 x 24 mL

RNase A

15 μL

160 μL

800 μL

Instruction Booklet

1

1

1

Buffer BL contains chaotropic salts that may form combustive compound with bleach. Use gloves and protective eyeware when handling this solution.

Before Starting                                                          

Please read the entire booklet to become familiar with the EZgeneTM Soil gDNA Kit protocol.

  •  Prepare Buffer LX stock solution by adding 10 μL β-mercaptoethanol per 1 mL Buffer LX before use. Each sample will require 600 μL of this solution.
  •  Preheat Buffer LX and Elution Buffer at 65 oC. Make sure the crystal in Buffer LX is completely dissolved.
  •  Dilute DNA Wash Buffer with absolute ethanol as follows and store at room temperature. Add 8 mL (GD2412-00) or 60 mL (GD2412-01) or 96 mL (GD2412-02) to each DNA Wash Buffer bottle before use.

Trouble Shooting Guide

Problem

Possible Reason

Suggested Improvement

A260/230 ratio is

low

inefficient elimination of inhibitory compounds

Repeat the DNA isolation with a new sample, be sure to mix the sample with DH reagent thoroughly and extract the sample with DH reagent twice.

Salt contamination

  • Repeat the DNA isolation with a new sample.
  • Make sure the column is dried before elution.
  • Wash the column with extra DNA Wash Buffer.

DNA Wash Buffer prepared

with lower percentage ethanol

prepare DNA Wash Buffer with 96-100% ethanol

A260/280 ratio is high

High RNA contamination

Be sure to treat the sample with RNase A. See step 5.

Low DNA yield or no DNA eluted

Sample stored incorrectly

Sample should be stores at -20 oC.

Poor homogenization of sample.

Repeat the DNA isolation with a new sample, be sure to mix the sample with Buffer LX and glass beads.

Use long beads beating time to make sure the sample are fully homogenized and cell are lysed.

Incorrect Buffer BL was added before loading to the column

Repeat the DNA isolation with a new sample

DNA washed off.

Dilute DNA Wash Buffer Concentrate by adding appropriate volume of absolute ethanol prior to use (page 3).

Problems in downstream applications

Ethanol residue in the elute

Be sure to completely dry the column before elution

Little or no supernatant after

initial centrifuge step

insufficient centrifugal force

Check the centrifugal force and increase the centrifugal time if necessary.

sample can not pass through the column

Clogging column

Check the centrifugal force and increase the time of centrifugation

Limited Use and Warranty

This product is warranted to perform as described in its labeling and in Biomiga's literature when used in accordance with instructions. No other warranties of any kind, express or implied, including, without limitation, implied warranties of merchantability or fitness for a particular purpose, are provided by Biomiga. Biomiga's sole obligation and purchaser's exclusive remedy for breach of this warranty shall be, at the option of Biomiga, to replace the products, Biomiga shall have no liability for any direct, indirect, consequential, or incidental damage arising out of the use, the results of use, or the inability to use it product.

For technical support or learn more product information, please contact us at (858) 603-3219 or visit our website at http://www.biomiga.com/

Operating Protocol

                 


  • Model: GD2412-01
  • Manufactured by: Biomiga

Additional Images