Are you still bothered by the low efficiency of homologous recombination of multi-fragment or super-long fragment and the small number of colonies?
Still for the carrier or fragment is more precious, low yield, in order to save raw materials, the use of small system low concentration ligation, but the ligation efficiency is low and bothered?
YEASEN product: Significantly improved, low concentration, small system and ultra-long fragments of the ligation efficiency, high positive cloning rate, help you to easily complete the ligation.
Comparison of one-step cloning with traditional enzyme-restriction cloning methods
Product Advantage
Orientation: Seamless assembly of 1-7 DNA fragments.
Simple: Multiple fragments can be ligated in one tube.
Fast: Fastest ligation in 5 min.
High efficiency: High positive cloning rate.
Small system: The total ligation system can be as low as 6 μL.
Overlength: Above 25 kb (Carrier+fragment length) for ligation.
Performance Display
- Efficient recombination of a single fragment 6 μL small system
Fig. 1 10 ng low-input single fragments were ligated in the non-resuscitation receptive cells state of the 6 μL small system. The results showed that 10923ES had better performance than competing products, with high ligation efficiency, more colonies and a positive rate of 100%. A-B: Reconstituted conversion plate. The molar ratio of carrier (10 kb) to insert fragment (1 kb) was 1:2. C: PCR identification of insert fragment. M: YEASEN 10505ES. Carrier volume: 40 ng/6 μL Reaction system, reaction conditions: 50℃, 15 min.
- Multi-fragment efficient recombination
Fig. 2 Six fragment (7.6 kb) ligated. The results showed that 10923ES had better performance than competing products, with more colonies and positive rate of 100%. A-B: Reconstituted conversion plate. The molar ratio of carrier (11.6 kb) to insert fragment (7.6 kb) was 1:2. C: PCR identification of insert fragment. M: YEASEN 10505ES, PCR length 2.6 kb. Carrier volume: 200 ng/20 μL Reaction system, reaction conditions: 50℃, 50 min.
- Low concentration multi fragment efficient recombination
Fig. 3 Ligate the low concentration five and six fragments separately. The results showed that 10923ES exhibited stable connectivity at low concentrations, with a high bacterial count and a 100% positivity rate. A-B: Recombinant Conversion Plate. C-D: Insertion fragment PCR identification electrophoresis image, M: YEASEN 10505ES, PCR length 2.6 kb.
- Reagent Stability
Fig. 4 After being placed at 37℃ for 7 days, the results showed that 10923ES remained stable in linking the five and six fragments at low concentrations. A-F: Recombinant Conversion Plate. G-H: Insertion fragment PCR identification electrophoresis image, M: YEASEN 10505ES, PCR length 2.6 kb.
Customer Case
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Efficient reorganization of small systems
Fig. 5 10923ES was ligated to a single fragment in a 10 μL small reaction system, with a positive rate of 100%. A: Reconstituted conversion plate. B: PCR identification of insert fragment. Carrier size: 5,631 bp; Target fragment size: 1,384 bp.
Fig. 6 10 μL Small system single fragment junction and point mutation clone, 100% positive rate. A & C: Recombination transformation plates. B: PCR identification of insert fragments, lanes 1-5 are colony 1, colony 2, negative control, positive control (genome as template), positive control (PCR purified fragment as template). D: Electrophoregram of PCR identification of insert fragments.
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Efficient recombination of low-concentration short fragments
Fig. 7 10923ES connecting the low concentration fragments. Compared with competing products, the number of colonies was high, and the positive rate (10 / 11) was> 90.9%. A-B: Reconstituted conversion plate. C: PCR identification of insert fragment. Carrier size: 3,000 bp; target fragment size: 441 bp.
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Efficient and fast multi segment ligation
Fig. 8 10923ES four segment ligation, high GC and short segment combination connection, compared to S brand, has a higher bacterial count of 10923ES and a positive rate of 100%. D: Recombinant conversion tablet. E: Insert fragment sequencing identification map. F: Sanger sequencing confirmed the successful connection between vector and fragments, and the sequencing at the connection site was correct.
Strength Star Product Recommendation
10923ES Order Channel
|
Product Orientation |
Name |
Art.No |
Specifications |
|
1 – 7 fragments were joined in one step and the recombination reaction was completed in the fastest 5 min |
Hieff CloneTM Universal II One Step Cloning Kit |
20 T/50 T |
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