A method for improving dna cloning and assembly efficiency by using short-chain primer suppression
A short-chain, purpose-built technology that can be used in DNA preparation, recombinant DNA technology, DNA/RNA fragments, etc., and can solve problems such as assembly efficiency that cannot be ignored
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Embodiment 1
[0125] Example 1. A method for improving DNA cloning and assembly efficiency by using short-chain primer suppression
[0126] The specific test process in this embodiment is as attached figure 2 shown.
[0127] 1. Preparation of 7 circular recombination vectors and receiving vector x
[0128] Circular recombination vector 1 includes DNA fragment A 1, DNA fragment 1 to be spliced, DNA fragment B 1 and backbone carrier from upstream (the nucleotide sequence of the backbone carrier is after removing nucleotides 4799 to 5663 in sequence 1 The obtained sequence); the DNA fragment A1 in the circular recombination vector 1 is the 23rd-55th position in the sequence 11, and the DNA fragment B1 is the 395th-426th position in the sequence 11;; the circular recombination vector 1 The nucleotide sequence of the DNA fragment A 1 that is identical or complementary to the short-chain primer A is the 30th-51st position in the sequence 11, the nucleotide sequence of the DNA fragment 1 to be ...
Embodiment 2
[0315] Example 2, the application of the method of short-chain primer suppression in the splicing of multiple fragments
[0316] The principle of this embodiment is attached Figure 4 shown.
[0317] 1. Preparation of 7 DNA fragments
[0318] The nucleotide sequence of DNA fragment 1 is shown as sequence 25 in the sequence table. DNA fragment 1 sequentially includes DNA fragment A1, DNA fragment 1 to be spliced and DNA fragment B1; DNA fragment A1 and short chain in DNA fragment 1 The nucleotide sequence identical or complementary to primer C is the 27th-53rd nucleotide molecule in sequence 25; the nucleotide sequence of DNA fragment 1 to be spliced is 54th-256th in sequence 25, DNA fragment B1 and The same or complementary nucleotide sequence of the short-chain primer D is the 257th-281th position in sequence 25;
[0319] The nucleotide sequence of DNA fragment 2 is shown as sequence 26 in the sequence table. DNA fragment 2 includes DNA fragment A 2, DNA fragment 2 to ...
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