Part:BBa_K4897001

BS DNA-162 (BS DNA 2.0) using in L-RCA for detecting P. acne

What is it?

BS DNA-162 was designed by BS United China as a single-stranded DNA segment complementary to the 131 base pairs of the 16S rRNA gene of P. acne. The composition of the DNA has three categories: binding region (two ends), amplification region, and random region. The binding region is the key element in reacting with the P. acne 16s rRNA gene [1]. The DNA ligase will perform the ligation of the single-stranded DNA meanwhile the phi29 will generate double-stranded DNA through amplification primers

Usage and Biology

Fig. 1. The process of BS DNA binding to P. acne DNA.

BS DNA-162 is the red line like a padlock. The black sequence represents the P. acne’s DNA specific to the binding. The red boxes represent the binding regions of the two ends of BS DNA-162 (notice that the two ends are not connected). The purple boxes represent the random sequences generated. The Blue box represents the amplification region for RCA.

BS DNA-162 would bind the bacterial DNA of P. acne and show great sensitivity and specificity.

Characterization

Fig.2. Amplification results of using BS DNA-162

Lane 1 is a 100 bp DNA ladder. Lane 2 is a positive control tested by synthetic and isolated P. acne DNA. Lane 3 is a positive result tested by samples from human faces through washing.

The positive control and the positive result both have amplification results beyond 162, the initial size, suggesting successful DNA ligation and amplification.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 83
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 57
  Illegal SpeI site found at 83
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 68
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 83
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 83
  
• 1000
  COMPATIBLE WITH RFC[1000]

Reference

[1]Nakamura, M., Kametani, I., Higaki, S., & Yamagishi, T. (2003). Identification of Propionibacterium acnes by polymerase chain reaction for amplification of 16S ribosomal RNA and lipase genes. Anaerobe, 9(1), 5–10. https://doi.org/10.1016/s1075-9964(03)00061-1.