DP336-TIANamp Soil DNA Kit-181121

TIANamp Soil
DNA Kit
For isolation of genomic DNA from soil
www.tiangen.com/en
This product is for scientific research use only. Do not use in
medicine, clinical treatment, food or cosmetics.
DP210831
TIANamp Soil DNA Kit

(Spin Column)
Cat. no. 4992288
Kit Contents
4992288
Contents
50 preps
Buffer SA 45 ml
Buffer SC 5 ml
Buffer HA 15 ml
Buffer HB 15 ml
Buffer GF 70 ml
Buffer PWS 15 ml
Buffer TE 15 ml
1 mm Grinding Beads 15 g
Spin Columns CB3 50
Collection Tubes 2 ml 50
Handbook 1
Storage
TIANamp Soil DNA Kit should be kept in dry place and can be stored at
room temperature (15-30°C) for up to 15 months. If any precipitate forms
in the buffers, it should be dissolved by warming the buffers at 37°C before
use.
TIANamp Soil DNA Kit Handbook

1
Introduction
TIANamp Soil DNA Kit uses a unique buffer system, by which the humic
acid in soil sample could be completely removed. 1 mm Grinding Beads
are also applied in this kit to process the lysis of components of soil sample
in order to guarantee the integrity of gDNA.
The genomic DNA isolated by this product is highly pure and has high
integrity, so it serves as an excellent template for downstream molecular
biological experiments like PCR analysis and restriction enzyme digestion.
Product Features
Wide application:
Good for gDNA isolation from flower bed soil, potting soil, farmland soil,
forest soil, sludge, red soil, black soil, dust and many other kinds of soil
samples.
Convenient operation:
The whole experimental procedure could be finished within a relatively
short time.
High purity:
With the application of spin column method, gDNA isolated by this kit have
high purity and is good for downstream experiments.
Important Notes (Please read before use)

1. Fresh soil sample could have a higher yield. Best storage conditions of
different soil samples should be checked before sampling.
2. All centrifugation steps should be carried out in a conventional table-
top microcentrifuge at room temperature (15-30°C).
3. Avoid pipetting precipitate in any supernatant collection steps, or else
the column would be blocked and the product purity would be affected.
4. Appropriate volume of ethanol need to be added to Buffer PWS as
indicated on the bottle tag before use.
5. Excessive amount of gDNA could inhibit the PCR reaction, so please
dilute the gDNA to an optimal concentration in such a circumstance.

2
Protocol
Ensure that Buffer PWS have been prepared with appropriate volume of ethanol
as indicated on the bottle tag and shake thoroughly.
1. Add 750 μl Buffer SA and 0.25 g Grinding Beads to a 2 ml microcentrifuge tube.
2. Add 0.25 g soil sample to the 2 ml microcentrifuge tube, mix by vortex for 15
sec.
3. Add 60 μl Buffer SC to the sample and mix by vortex for 10 min.
Note: Check if there is any precipitate in Buffer SC before use, if there is,
please dissolve the precipitate by warming at 37°C.
4. Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant
(around 500 μl) to a new 2 ml microcentrifuge tube.
5. Add 250 μl Buffer HA, mix by vortex for 5 sec, and incubate the tube at 4°C for
5 min.
6. Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant to a
new 2 ml microcentrifuge tube, add 200 μl Buffer HB and mix, incubate the
tube at 4°C for 5 min.
Note: Avoid pipetting precipitate when transferring supernatant, or else the
purity of product would be affected.
7. Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant to a
new 2 ml microcentrifuge tube, add 1,200 μl Buffer GF and mix by inverting.
Note: Avoid pipetting precipitate when transferring supernatant, or else the
purity of product would be affected.
8. Transfer 700 μl solution from step 7 to a Spin Column CB3 (place the column in
a collection tube), centrifuge at 12,000 rpm (~13,400 × g) for 30 sec, discard
the flow-through, put the column back to the collection tube.
Note: The capacity of Spin Column CB3 is 700 μl, if the sample volume
exceeds 700 ul, centrifuge successive aliquots in the same column. Discard
the flow-through after each centrifugation.
9. Add 500 μl Buffer PWS (Ensure that ethanol (96-100%) has been added)
to the Spin Column CB3, centrifuge at 12,000 rpm (~13,400 × g) for 30 sec,
discard the flow-through, put the column back to the collection tube.
10.Add 500 μl 70% ethanol to the Spin Column CB3, centrifuge at 12,000 rpm
(~13,400 × g) for 30 sec, discard the flow-through, put the columnback to the
collection tube.
11.Centrifuge at 12,000 rpm (~13,400 × g) for 2 min, discard the flowthrough.
12.Incubate the Spin Column CB3 at room temperature (15-30°C) for several
minutes to completely dry the residual washing buffer in the column. Place the
Spin Column CB3 in a new clean microcentrifuge tube, and pipet 50-100 μl
Buffer TE directly to the center of the membrane. Incubate at room

3
temperature for 2-5 min, and then centrifuge for 2 min at
12,000 rpm (~13,400 × g), collect the flowthrough to the microcentrifuge
tube.
Note: In order to get a high yield, flow-through could be pipetted
back to the Spin Column CB3 and incubated at room temperature
for 2 min, then centrifuge again at 12,000 rpm (~13,400× g) for 2
min. The pH value of elution buffer has a great impact on elution
efficiency, we suggest that the pH value should be within the range of
7.0-8.5 if distilled water is used as elution buffer. Low pH value (pH<7)
would significantly reduce the efficiency of elution. DNA product
should be stored at -20°C to avoid the degradation.
Analysis of DNA concentration and purity

Size of gDNA extracted by this kit is related with the sample storage
condition, shearing force during operation and some other factors. Purified
DNA can be analysis by electrophoresis gel and UV-Spectrophotometer.
DNA has a significant peak at OD260. An OD260 of 1 corresponds to a 50
μg/ml of dsDNA solution or a 40 μg/ml of ssDNA solution.
OD260/280 ratio value should be within 1.7-1.9. If ddH2O is used to elute
DNA, the ratio value would be lower, since the pH value and the existence
of ion could affect the absorption value.

4

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TIANamp Soil

TIANamp Soil DNA Kit

Cat. no. 4992288

TIANamp Soil DNA Kit Handbook

Important Notes (Please read before use)

TIANamp Soil DNA Kit Handbook

TIANamp Soil DNA Kit Handbook

Analysis of DNA concentration and purity

TIANamp Soil DNA Kit Handbook

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