Hi all,
I have analyzed a total of 7 samples coming from the bacteriophage fraction of human faeces. Purification included resuspension of the sample in buffer SM and homogenized sample was then filtered by low protein absorption filters. Afterwards samples were DNAse treated in order to eliminate non viral DNA or non capsided DNA. Also, samples were treated with chloroform to eliminate membrane vesicles containing DNA. Then, posterior membrane filters allowed to eliminate another residual bacteria.
Samples were sequenced in an Illumina Miseq paired end 2x150 bp. Samples were de novo assembled and taxonomically classified.
The problem is that a high proportion of bacterial DNA was detected in my samples so I wonder if someone could know what happened.
1) Could it be possible that bacterial DNA has been integrated in bacteriophages?
2) Should purification have included any other step?
Any other clue would be appreciated
I have analyzed a total of 7 samples coming from the bacteriophage fraction of human faeces. Purification included resuspension of the sample in buffer SM and homogenized sample was then filtered by low protein absorption filters. Afterwards samples were DNAse treated in order to eliminate non viral DNA or non capsided DNA. Also, samples were treated with chloroform to eliminate membrane vesicles containing DNA. Then, posterior membrane filters allowed to eliminate another residual bacteria.
Samples were sequenced in an Illumina Miseq paired end 2x150 bp. Samples were de novo assembled and taxonomically classified.
The problem is that a high proportion of bacterial DNA was detected in my samples so I wonder if someone could know what happened.
1) Could it be possible that bacterial DNA has been integrated in bacteriophages?
2) Should purification have included any other step?
Any other clue would be appreciated
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