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Product Highlight
HMW Genomic DNA Purification Kit ............................. ........................................................
Catalog Number P4001A/P4001B
For purification of high molecular weight genomic DNA from animal whole blood, cultured cells, animal tissues, plant tissues, bacteria, yeast, fungi.
HMW genomic DNA purification kit provides a simple, efficient and reliable method for isolating genomic DNA with high molecular weight from varieties of organism. Although the kit was designed for processing from small volume of starting material, it is easy to scale up to larger volume of samples. The procedure provided by the kit eliminates many disadvantages, such as time-consuming methods, toxic components, low yield, as compared to other commercial kits.
The purified RNA is immediately ready to use in various downstream applications, such as PCR, real-time PCR, enzymatic digestion, blotting, and hybridizations.
Key Features
- Easy processing: only 3 steps(lysis-protein removal- DNA precipitation) are needed for most samples
- The molecular weight of purified DNA is 100-150kb.
- High yield and purity: OD 260/OD 280 of purified DNA is between 1.8-2.0.
- Safety for handling, shipping and storage: No phenol/chloroform extraction, no toxic chaotropic salts.
- No column or resin is needed.
- Extracted DNA is ready for downstream applications.
Kit Contents
Catalog Number |
P4001A |
P4001B |
Kit Size |
100 preps |
200 preps |
Buffer RL |
50 |
100 |
Lysis Buffer LB |
35 ml |
70 ml |
Buffer SN |
15 ml |
30 ml |
Buffer TE |
20 ml |
20 ml |
RNase A solution |
0.25 ml |
0.5 ml |
Handbook |
1 |
1 |
Genomic DNA yields from various starting materials
Tab.1 Examples of purified genomic DNA from various starting materials
Starting material |
Amount |
Typical gDNA yield(~µg) |
Human |
Whole blood |
300µl |
20 |
Mouse blood and tissues |
Whole blood |
300µl |
15 |
Spleen |
20mg |
30 |
Liver |
20mg |
30 |
Lung |
20mg |
10 |
heart |
20mg |
10 |
Kidney |
20mg |
30 |
Tail |
1cm |
30 |
Cultured cell lines |
HCT-116(human) |
2x10 6 |
30 |
SW-620 (human) |
2x10 6 |
30 |
MCF7(human) |
2x10 6 |
30 |
Hela(human) |
2x10 6 |
30 |
CHO(hamster) |
2x10 6 |
30 |
E. coli |
Strain K-12 |
1ml(~2x10 9) |
30 |
Plant |
Bean leaf |
30mg |
15 |
Yeast |
Pichia pastoris |
1ml(~2x10 8) |
10 |
Example: Protocol for genomic DNA purification from 2 x 10 6 of cultured cells
Step 1: Lysis of cultured cells
- Transfer the solution containing cultured cells into a 1.5 ml tube. Centrifuge at top speed for 30 seconds to pellet the cells.
- Decant the supernatant. Pulse-vortex the tube for 5-10 seconds to resuspend the pellet in the residual liquid.
- Add 350 μl of lysis buffer LB. Pipett up and down to resuspend the pellet completely.
- Add 5 µl of RNase A solution. Mix thoroughly by vortexing briefly. Incubate the mixture at 37ºC for 30 minutes.
Step 2: Removal of cell debris and proteins
- Add 150 µl of buffer SN . mix thoroughly by vortexing for 10 seconds.
- Centrifuge at maximum speed for 3 minutes. Decant gently the supernatant into a new tube.
Step 3: Isolation of genomic DNA
- Add I ml of ethanol into the tube. Invert the tube 4-5 times to mix the solution thoroughly
- Centrifuge at maximum speed for 2 minutes at room temperature.
- Add 1 ml of 70% ethanol, Gently invert the tube 4-5 times. Decant the supernatant carefully.
- Repeat the wash step( step 3 ) 2 times.
- Stand the tube with cap open for 15 minutes at room temperature.
- Add 100 μl of buffer TE. Dissolve the pellet by Pipetting up and down. Incubate the tube at 60ºC for 1-2 hours.
- The DNA is ready for use.
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