Animal experiment is essential in biomedical research. It contains establishment of a nude mouse tumor model and other disease model. Besides, gene transfection in vivo is included as well. As previous reported, gene delivery targeting to specific organs can be achieved through AAV (Adeno-associated virus). But how to achieve virus injection in specific organs Three […]
Category Archives: protocol
Genomic DNA primer design 1 Design genomic DNA primers that are approximatley 18 to 22 basepairs in length and have 45-55% GC content. For best results, use primers with a Tm greater than 55 C. Design primers to yield amplicon length between 400-500 bp. In addition, design primers so that the predicted cleavage site is not […]
Lentivirus Protocol Download Lentivirus Production Protocol–Genemedi 1. Lentivirus plasmid construction The gene of interest is cloned into one of the LTR/MCS-containing lentivirus vectors to generate pLV-GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration. 2. Lentivirus packaging The recombinant lentivirus viral plasmid pLV-GOI is co-transfected into the 293T with envelope […]
Lentivirus Protocol Download This protocol is for the stable cell line construction based on puromycin selection. Day 1: Seed target cells in 24-well plates. The number of seeding cells differs according to the cell proliferation rate.Day 2: Target cells should be approximately 50%-70% confluent. For polybrene accessible cells, mix the culture medium with proper concentrations of polybrene. […]
Adenovirus protocol Download Adenovirus Production Protocol–Genemedi 1. Adenovirus plasmid construction The gene of interest is cloned into one of the ITR/MCS-containing adenovirus vectors to generate pAd -GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration. 2. Adenovirus packaging The recombinant adenovirus viral plasmid pAd-GOI is co-transfected into the 293A with […]
AAV protocol Download AAV Production Protocol–Genemedi 1. AAV plasmid construction The gene of interest is cloned into one of the ITR/MCS-containing AAV vectors to generate AAV-GOI. The purity and RNAse contaminants of AAV viral plasmid should be taken into consideration. 2. AAV packaging The recombinant AAV viral plasmid AAV-GOI is co-transfected into the AAV-293 cells […]
CRISPR/Cas9 protocol Download Gene editing, also called genome editing, is a group of technologies to change the sequence of DNA in the genome. Several approaches to genome editing have been developed, including Zinc Finger, TALEN, and CRISPR/Cas9. Compared with the other gene editing tools, CRISPR/Cas9 system is faster, cheaper, more accurate, and more efficient, showing […]
Protocol Download 1. To prepare the AuNP (gold nanoparticle) conjugate, the recombinant protein dissolved in PBS (1 mg/mL) was added to the mixture of 1mL AuNP colloid (40nm in diameter, OD=1) and 0.1mL of borate buffer (0.1 M, pH 8.5). Incubation for 30 min at room temperature. 2. 0.1mL of 10mg/mL BSA in PBS was […]
Protocol Download Standard biopanning 1. S-RBD was diluted with 0.05 M PH 9.6 carbonate buffer [1]. Then add 100 μL to each well of 96-well Maxisorp plates at 4°C overnight. 2. Aspirate the solution in the well and wash each well three times with Wash Buffer [2] (300 μL/well). Complete removal of liquid at each step is essential […]
Protocol Download 1. The Capture Antibody #1 was diluted to a concentration of 1 ~ 10 ug/ml with 0.05 M PH 9.6 carbonate buffer [1]. Then add 100 μL to each well of polystyrene plate at 4℃ overnight. 2. Aspirate the solution in the well and wash each well three times with Wash Buffer [2] (300 μL/well). Complete removal of […]
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