A mask with excellent moisturizing and repairing effects
A facial mask and membrane-penetrating peptide technology, which is applied to medical preparations containing active ingredients, skin care preparations, and cosmetic preparations, etc., can solve problems such as high cost, difficult mass production of EGF, and difficulty in fully exerting its efficacy. To achieve the effect of enhancing solubility
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Embodiment 1
[0033] Example 1: Acquisition of target DNA
[0034] (1) The gene sequence of the target gene is:
[0035] Using 5'-GCAAGCGCATTGCACATGTACGGCTATAGCAGATA-3' (SEQ ID NO: 1) with the target gene transcribed messenger RNA as a template, reverse transcribed into complementary single-stranded DNA, and then synthesize double-stranded DNA under the action of T4DNA ligase, that is, the purpose Gene.
[0036] (2) PCR amplification
[0037] Denaturation: high temperature dissociates double-stranded DNA into single-stranded DNA (94°C, 30s);
[0038] Annealing: the primer (pACT2 GAL4 AD pACT2-R) binds to the complementary region of the template DNA at low temperature to form a hybrid molecule (55°C, 30s);
[0039] Extension: mesophilic extension, in DNA polymerase, dNTP, Mg 2+ In the presence of DNA polymerase, DNA polymerase catalyzes the extension of the DNA strand starting from the primer from 5' to 3' to synthesize a DNA daughter strand complementary to the template DNA strand (70-7...
Embodiment 2
[0041] Example 2: Electrophoresis detection and recovery and purification
[0042] (1) Electrophoresis
[0043] ① Prepare an appropriate amount of buffer for electrophoresis and gel preparation (1X TAE Buffer)
[0044]②Accurately weigh the agarose powder (1g) according to the amount of gel and gel concentration, and add it to an appropriate conical flask.
[0045] ③ Add a certain amount of electrophoresis buffer (1X TAE Buffer 100ml).
[0046] ④ After melting is complete, cool to about 60℃, add EB5-7ul, and mix well.
[0047] ⑤Pour the solution into the plastic mold, before inserting a comb in the appropriate position.
[0048] ⑥It will solidify at room temperature. If it is not used immediately, wrap the gel with plastic wrap and store it at 4℃.
[0049] Mix an appropriate amount of PCR product with an appropriate amount of bromophenol blue, add it to the gel tank, and add an appropriate amount of DNAMaker to the right tank to start electrophoresis. Observing the electro...
Embodiment 3
[0055] Example 3: Construction of recombinant expression vector
[0056] The selected recombinant expression vector plasmid vector is pBR322, and the construction of the recombinant plasmid includes the following steps:
[0057] (1) Since one of the parents of the pBR322 plasmid is the pMB1 plasmid, firstly, based on pMB1, the Tn3 translocation of the Rldrd19 plasmid was introduced to obtain a 13.3kb pMB3 plasmid;
[0058] (2) The molecular weight of pMB3 obviously makes it unsuitable as a carrier, so most of the useless fragments must be lost by digestion under the conditions of EcoRI activity, and the sticky ends of the remaining small fragments of DNA are ligated and formed. pMB8 plasmid (2.6kb);
[0059] (3) At this time, another pSC101 plasmid was digested under EcoRI activity conditions to produce a tetr-resistant DNA fragment, which was integrated with pMB8 to form pMB9 plasmid (5.3kb). At this time, pMB9 is ampstetr phenotype;
[0060] (4) pMB9 has preliminarily fun...
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