Construction method and application of microbial diversity library
A library construction and microbial technology, applied in biochemical equipment and methods, microbial measurement/testing, chemical libraries, etc., can solve the problems of sample abundance errors, inability to construct diverse libraries, inability to identify multiple species in the same sample, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0186] 1. Synthesis of special index primer adapter:
[0187] The special index primer adapter is a double-stranded adapter with a sticky end, one end of the special index primer adapter is a first sticky end, and the first sticky end includes at least a special end sequence, and the special end The sequence is used to identify the target region of the microbial genome DNA, and is connected to the single-stranded DNA of the target region through complementary pairing. The target region may be 16S rRNA region, 18S rRNA region or ITS region, etc. The structure of the special end sequence is "optional sequence-recognition target sequence", the optional sequence is used to adjust the Tm of the linker and ensure that the Tm between the linkers is similar, and the recognition target sequence is used for matching with the target region Complementary pairing of nucleotide sequences.
[0188] 1. As an example, use Primer1 and Primer2 to synthesize special index primer adapters,
[018...
Embodiment 2
[0340] (1) Take 10 ng of feces samples and divide them into 6 parts on average, and group them according to Table 14 below. Negative controls were set for each group, named N1&N2 respectively:
[0341] Table 14
[0342]
[0343] Among them, A-C samples were extracted and quantified using picogreen double-strand quantitative dye, and the quantitative results were all lower than the detection limit.
[0344] The general database construction scheme is as follows:
[0345] 1. Prepare the following PCR reaction mixture:
[0346] Table 15
[0347] sample
Volume (ul)
KAPA HiFi HotStart ReadyMix (2x)
25
Primer library (10μM)
2
NF water
23
total capacity
50
[0348] Wherein, the primer library refers to primer pairs well-known to those skilled in the art for amplifying microorganisms such as bacteria, fungi, archaea, denitrifying bacteria, nitrogen-fixing bacteria, nitrite reducing bacteria, ammonia oxidizing bacteria, a...
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, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com