To start with from SDS Gel,

Dear Aadarsh,

1. Alignment to the well is crucial, I can see some angle from the base/well position.

2. Many bands in each column, marker type?RAPD's, SSR's?specificity?

3. How long and at what voltage the gel runs? it seems the gel need to run more time

   to see if we get a specific band.

Next about the spore germination,

1. required material: microscope, slides, films, sample

2. drop a sample on the slide, cover with film, and see through Microscope if you see germination

3. repeat step 2 for two three days until you see germination on your scope

4. transfer to petridisch with PDB media

5. place them to grow with light on the shelf

6. transfer them to liquid media with in 4 to 6 weeks, depending on the nature of the fungus

7. Extract DNA and save the sample to silica saves for storage

8. perform other molecular works: PCR, sequencing, .....

Good luck Aadarsh.

Dear Aadarsh,

You could use dilution and plating for microbial enumeration (in this case spores).

For that, prepare your fungal suspension with the usual concentration (10e7). Then make tenfold serially diluted suspensions in NaCl (0,85%) up to 10e2 spores/ml.

Transfer 100µL from each one of the last dilutions (10e2, 10e3 and 10e4 spores/ml), from the less to the more concentrated dilution into PDA. Then spread the diluted suspension over the surface of the agar plate.

After incubation you count the number of colonies in each plate. If you get about 10 (5-20) colonies for the dilution 10e2, 100 colonies (50 - 200) for 10e2 and 1000 colonies ( all the plate is full) for 10e3, then the viability of your spores (and therefore germination) is OK and maybe there is another reason that is inhibiting the production of cellulases.

Do you get the same growth with the two inoculation methods?

The Best,

Diana

Fungal spores sometimes need both free water (not water vapor) and nutrients and sometimes even a hard surface for germination.