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Old 11-25-2013, 08:14 PM   #1
lebonahd
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Default Hai, I need clear protocol for DNA Barcoding in bacteria

DNA Barcoding in bacteria
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Old 12-17-2013, 08:45 AM   #2
cement_head
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Try starting with this website: http://www.earthmicrobiome.org/emp-s...protocols/16s/
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Old 12-17-2013, 11:57 PM   #3
lebonahd
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Default Barcoded pyrosequencing

Thank you sir for your reply, I'll surely follow that link. But I have a doubt, please clarify me.
Can Barcoded pyrosequencing identify bacteria at species level? Because I want to identify bacteria found in human milk at species level by Barcoded pyrosequencing. So, please suggest me sir.

Thank you,
D.E. Lebonah
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Old 12-18-2013, 02:02 AM   #4
cement_head
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Quote:
Originally Posted by lebonahd View Post
Thank you sir for your reply, I'll surely follow that link. But I have a doubt, please clarify me.
Can Barcoded pyrosequencing identify bacteria at species level? Because I want to identify bacteria found in human milk at species level by Barcoded pyrosequencing. So, please suggest me sir.

Thank you,
D.E. Lebonah
You should be able to identify bacteria at the species level. The barcoding (indexing) will be used to identify the sample source (which individual person, which aliquot) and will not have an influence on the taxonomy. It is the variability in the 16S region that will be the determining factor in attributing species level taxonomy to the obtained sequence data.

By the way, the protocol I sent you an URL to is for Illumina (Solexa) sequencing, not Roche's Pyrosequening.

Regards,
CH
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