Damage-free cold extraction method for genome DNA of small insects
A genomic and cold extraction technology, applied in the field of molecular biology, can solve the problems of insufficient DNA release, genomic DNA degradation, affecting DNA quality, etc., to reduce DNA degradation, enhance digestion, and improve yield.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0028] Example 1
[0029] Extract the genomic DNA of Hymenoptera ants (sample 1).
[0030] 1 Preparation of extraction buffer
[0031] Prepare 400 μL extraction buffer: 40 μL 10×PCR Buffer II, 1.8 μL NP40, 1.8 μL Tween20, 12 μL proteinase K (20 mg / mL), 344.4 μL ddH 2 O.
[0032] 2 Pretreatment of worms
[0033] First use 95% alcohol to carefully clean the impurities on the insect's body surface, then wash it with distilled water 3 times, and finally use absorbent paper to absorb the surface water of the insect body.
[0034] 3 Extraction of genomic DNA
[0035] (1) Put the ants in a centrifuge tube, add 300 μL of extraction buffer, bath at 50°C for 1 h, and gently invert and mix once every 15 min;
[0036] (2) Place at -20°C for 15 hours, cold extract genomic DNA;
[0037] (3) Add 9 μL of proteinase K (20 mg / mL), in a water bath at 50°C for 1 h, gently invert and mix once every 15 minutes;
[0038] (4) Water bath at 95℃ for 10 min to inactivate proteinase K;
[0039] (5) Take the ants out of...
Example Embodiment
[0045] Example 2
[0046] Extract the genomic DNA of Homoptera leafhopper (sample 2).
[0047] 1 Preparation of extraction buffer
[0048] Prepare 300 μL extraction buffer: 30 μL 10×PCR Buffer II, 1.35 μL NP40, 1.35 μL Tween20, 9 μL proteinase K (20 mg / mL), 258.3 μL ddH 2 O.
[0049] 2 Pretreatment of insect body (same as Example 1)
[0050] 3 Extraction of genomic DNA
[0051] (1) Put the leafhoppers in a centrifuge tube, add 200 μL extraction buffer, and bath at 50°C for 1 h, gently invert and mix once every 15 minutes;
[0052] (2) Place at -20°C for 15 hours, cold extract genomic DNA;
[0053] (3) Add 6 μL of proteinase K (20 mg / mL), in a water bath at 50°C for 1 h, gently invert and mix once every 15 min;
[0054] (4) Water bath at 95℃ for 10 min to inactivate proteinase K;
[0055] (5) Take the leafhopper out of the centrifuge tube with sterile tweezers, and put it in 95% alcohol for storage. The solution in the centrifuge tube contains the genomic DNA of the leafhopper, and it shou...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap