My supervisor has instructed me to conduct colony PCR on B. subtilis 6633, however, I am skeptical about it because colony PCR is commonly used to determine if a vector is present, I don't think that colony PCR is able to extract gDNA and amplify the amylase gene within the gNDA. Furthermore, I've heard that colony PCR is typically used for Gram-negative bacteria due to its thinner cell wall.
Irka Nur Adriana Binte Irwan Colony PCR has been done for Bacillus and other Gram+ bacteria and even for yeast.
See for reference:
Jagtap SC, Jagtap CB, Kumar P, Srivastava RB. Detection of Bacillus sphaericus mosquitocidal toxin genes by multiplex colony PCR. Canadian journal of microbiology. 2009 Feb;55(2):207-9.
Kumpfmüller J, Kabisch J, Schweder T. An optimized technique for rapid genome modifications of Bacillus subtilis. Journal of microbiological methods. 2013 Dec 1;95(3):350-2.
Afzal-Javan F, Mobini-Dehkordi M. Amplification, sequencing and cloning of Iranian native Bacillus subtilis alpha-amylase gene in Saccharomyces cerevisiae. Jundishapur Journal of Microbiology. 2013 Oct 1;6(8):1Q.
Colony PCR can be done for Gram-Positive Bacteria. I have done colony PCR for around 10 Gram-Positive bacteria.
Due to heating at 90 degrees in the water bath for 10 min, the cell wall will break easily and the compounds inside the cell will ooze out. TE buffer holds the DNA and the rest of the debris can be removed in centrifugation.
Instead of TE buffer, you can use DD water but the quality of DNA may decrease.
Yes, You can be done for Gram positive bacteria for good results
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