Prix FOB
Obtenir le dernier prix|
1 Set Minimum Order
Pays:
China
N ° de modèle:
EE101
Prix FOB:
Localité:
-
Prix de commande minimale:
-
Commande minimale:
1 Set
Packaging Detail:
EE101-01, 50rxns; EE101-02, 200rxns
Heure de livraison:
-
Capacité de Fournir:
1000 Set per Week
Payment Type:
T/T, L/C, D/A, D/P, Western Union, Money Gram, PayPal
Groupe de produits :
Personne àcontacter Dan
Dongsheng Technology Park, Haidian District, Beijing, Beijing
Cat.No. Â EE**1
Ordering Information:
|
EE*****1 |
*0 rxns |
|
EE*****2 |
**0 rxns |
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Storage:RNase A and Proteinase
K solutions at **0°C for one year; other components at room
temperature (****5°C) for one year.
Description
EasyPure® Genomic DNA Kit provides a simple and convenient way to isolate high quality genomic DNA from a variety of mammalian cells, tissues, E.coli and yeast. Cells and tissues are enzymatically lysed. DNA is bound to silica-based column. The isolated DNA is suitable for PCR, restriction enzyme digestion and southern blotting.
• DNA yield up to *5 μg.
• Complete removal of contaminants and inhibitors.
• DNA range of ****0 kb.
• Column based purification, no organic extraction or ethanol precipitation. Â
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Kit Contents
|
Component |
EE*****1 (*0 rxns) |
EE*****2 (**0 rxns) |
|
Lysis Buffer 2(LB2) |
6Â ml |
*4 ml |
|
Binding Buffer 2 (BB2) |
*8Â ml |
**0 ml |
|
Clean Buffer 2 (CB2) |
*5Â ml |
2×**0 ml |
|
Wash Buffer 2 (WB2) |
*2Â ml |
2×*2 ml |
|
Elution Buffer (EB) |
*5 ml |
*0 ml |
|
RNase A(*0 mg/ml) |
1 ml |
4×1 ml |
|
Proteinase K(*0 mg/ml) |
1 ml |
4×1 ml |
|
Spin Column with collection Tubes |
*0 each |
2×**0 each |
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Procedures
Before starting, adding appropriate volume of *****0% ethanol to WB2.
|
WB2 |
*2 ml |
2×*2 ml |
|
Ethanol |
*8 ml |
2×*8 ml |
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1. Preparing materials
• Mammalian Cells
a) Adherent cells: Remove the culture media from culture plate and harvest cells by trypisin or other methods. Collect cells by centrifuging at **0×g for 5 minutes, and then remove the supernatant .
b) Suspension cells: Harvest cells by centrifuging at **0×g for 5 minutes. Remove the supernatant.
c) Add **0 μl LB2 to the cell pellet, mix thoroughly by vortexing or pipetting.
Optional: If RNA-free genomic DNA is required, add *0 μl RNase A to the lysate.
d) Add *0 μl Proteinase K to the lysate. Mix the tube briefly by vortexing, then incubate at room temperature for 2 minutes.
• Mammalian Tissues (Prepare *5°C water bath or heater before starting).
a) Transfer ≤*5 mg (spleen≤*0 mg) chopped tissue to a sterile 1.5 ml microcentrifuge tube.
b) Add **0 μl LB2 and *0 μl Proteinase K to the tube. Make sure that the tissue is completely immersed in the tube.
c) Incubate at *5°C until the sample is completely lysed (3 hours are needed for most tissues; **8 hours or longer are needed for mouse tail; mix the lysate 2~3 times every hour).
Optional: If RNA-free genomic DNA is required, add *0 μl RNase A to the lysate, incubate at room temperature for 2 minutes.
d) Centrifuge at *2,**0×g for 5 minutes, transfer the supernatant to a sterile 1.5 ml microcentrifuge tube.
• E.coli Cells (Prepare *5°C water bath or heater before starting).
a) Transfer 1~5 ml cell culture (≤2×**9) to a 1.5 ml tube and centrifuge the tube at *2,**0×g for 1 minute. Discard the supernatant completely.
b) Add **0 μl LB2 and *0 μl Proteinase K into the tube. Resuspend the cell pellet by vortexing or pipetting. c) Incubate at *5°C for *5 minutes.
Optional: If RNA-free genomic DNA is required, add *0 μl RNase A to the sample, incubate at room temperature for 2 minutes.
 • Yeast Cells (Prepare *7°C, *5°C water bath or heater before starting)
Prepare fresh D-glucitol buffer (1 M sorbitol, *0 mM EDTA, *4 mM β-mercaptoethanol). Prepare lyticase. a) Harvest yeast cells (≤5×**7 ) by centrifuging at *2,**0×g for 1 minute. Discard the supernatant.
b) Add **0 μl D-glucitol buffer, *5 units lyticase to the pellet. Mix thoroughly and incubate at *7°C for 1 hour.
c) Centrifuge at 5,**0×g for *0 minutes. Discard the supernatant.
d) Resuspend pellets in **0 μl LB2 and *0 μl Proteinase K, mix thoroughly by votexing.
e) Incubate at *5°C for *5 minutes. Optional: If RNA-free total DNA is required, add *0 μl RNase A to the lysate, incubate at room temperature for 2 minutes.
f) Centrifuge at *2,**0×g for 5 minutes and transfer the supernatant to a sterile 1.5 ml microcentrifuge tube. 2. Add **0 μl BB2, mix by vortexing for 5 seconds, incubate at room temperature for *0 minutes.
3. Centrifuge the tube briefly and transfer all the lysate to a spin column. Centrifuge at *2,**0×g for *0 seconds. Discard the flow through.
4. Add **0 μl CB2, Centrifuge at *2,**0×g for *0 seconds. Discard the flow through.
5. Repeat step 4. 6
. Add **0 μl WB2 (check to ensure you have added ethanol) and centrifuge at *2,**0×g for *0 seconds. Discard the flow through.
7. Repeat step 6.
8. Centrifuge the empty column at maximum speed (≥*2,**0×g) for 2 minutes to remove residual WB2.
9. Place the spin column in a sterile 1.5 ml microcentrifuge tube. Add *****0 μl of Elution Buffer (preheated to *5°C) or sterile, distilled water (pH >7.0, preheated to *5°C) to the column matrix. Incubate at room temperature for 1 minute. Centrifuge at *2,**0×g for 1 minute to elute the isolated genomic DNA. Optional: To get more DNA by repeating step 9.
*0. Store the isolated DNA at **0°C.
Notes
• Do not use too many starting materials in case it affects the extraction performance
• Cut the tissue as small pieces as possible, in case it affects lysis. After complete lysis, the lysate presents sticky appearance, rather than gelatinous appearance.
• To ensure the quality of extracted DNA, use fresh material and avoid repeated freezing and thawing . DNA quality depends on the types of material and storage time.
• Use sterile tubes and pipette tips to avoid the contamination from DNase.
• You may perform the second elution step using the same microcentrifuge tube or different tubes.
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CITATIONS
Geng J, et al. ***3. Efficient Attenuation of NK Cell–Mediated Liver Injury through Genetically Manipulating Multiple Immunogenes by Using a Liver-Directed Vector. **0(9):*****9. J Immunol. IF=5.*2. PMID: *******2.
Xie K, et al. ***3. OxyR, an important oxidative stress regulator to phenazines production and hydrogen peroxide resistance in Pseudomonas chlororaphis GP*2. **8(*0):******3. Microbiol Res. IF=1.**3. PMID: *******5.
Liu Q. et al. ***2. Identification of the bacteriocin subtilosin A and loss of purL results in its high-level production in Bacillus amyloliquefaciens.. **3(**7):****8. IF= 2.**3. PMID: *******3.
Xu X. et al. ***2.Hypoxia induces downregulation of soluble guanylyl cyclase β1 by miR**4c*5p. **3(**7):****8. IF= 6.**1. PMID: *******7.
| Pays: | China |
| N ° de modèle: | EE101 |
| Prix FOB: | Obtenir le dernier prix |
| Localité: | - |
| Prix de commande minimale: | - |
| Commande minimale: | 1 Set |
| Packaging Detail: | EE101-01, 50rxns; EE101-02, 200rxns |
| Heure de livraison: | - |
| Capacité de Fournir: | 1000 Set per Week |
| Payment Type: | T/T, L/C, D/A, D/P, Western Union, Money Gram, PayPal |
| Groupe de produits : | Nucleic Acid Purification |
