Naked eye screening of recombinant baculovirus assay
Naked eye screening of recombinant baculovirus assay
The baculoviruses are a class of DNA viruses that exclusively infect arthropods in nature. The virus particles are rod-shaped, the genome is a double-stranded circular DNA molecule, and the DNA is compressed and packaged in the form of superhelix in the rod-like capsid, with the size ranging from 90 to 180 Kb. Currently, baculoviruses are widely used in pest control as highly efficient and safe pollution-free biological insecticides.
Operation method
basic program
Materials and Instruments
baculovirus Move Screening under a dissecting microscope1. invert the etching petri dish so that it is against the black background of the microscope and shine a bright light into the dish at an acute angle. For more product details, please visit Aladdin Scientific website.
Agarose
Dissecting microscope Inverted microscope
2. examine wild-type virus-infected cells for etchings containing inclusion bodies that appear refractive and have a slight yellow coloration.
3. examine the etchings of β-gal recombinant virus-infected cells that do not contain inclusion bodies and are gray in color.
4. circle the area in the dish that is covered with etchings and examine the circled etchings under an inverted microscope at a magnification of 400x to exercise the ability to distinguish the morphological subtleties of each etching with the naked eye.
Screening with an inverted light microscope1. Raise the right side of the etching dish slightly upward and examine each type of etch at 30x and 40x magnification. 2.
2. Recombinant etchings have an orange-red color, while wild-type etchings have a dark brown appearance due to aggregation of inclusion bodies. 3.
3. when the etch is seen under the microscope, spot the area under the etch on the petri dish and remove the dish from the microscope to enclose the etch. 4. invert the petri dish and place it on the microscope.
4. Place the dish upside down on the microscope stage and observe the etch at 400x magnification.Screening directly with the naked eye1. if the etchings are well formed, it is possible to recognize wild-type etchings from reconstituted etchings by holding the petri dish over your head and looking directly at the bottom of the dish.
2. the area around the wild-type etchings looks grayish-white, while the reconstituted etchings do not.3. after identifying the presumed recombinant viral etchings by either of the above methods, pick a few etchings, place the agarose block in 1 ml of serum-free medium, mix by oscillation on a vortex mixer, and the viral solution can be stored at 4°C for several months for backup.