Primer

Part:BBa_K4347000

Designed by: Victor Di Donato, Nicoletta de Maat   Group: iGEM22_Queens_Canada   (2022-07-07)
Revision as of 20:29, 21 July 2022 by Victor5688 (Talk | contribs) (→‎References)

Forward Inner Primer (FIP)

The forward inner primer (FIP) is one of the six primers needed to carry out Loop Mediated Isothermal Amplification (LAMP).

Usage and Biology

The loop-mediated isothermal amplification (LAMP) is a rapid method that allows for the specific DNA amplification through the utilization of multiple primers and three major steps[1]. This method relies on 3 major components- Bst, a DNA polymerase, and two sets of primers, and the target DNA/RNA. The first step, referred to as the “starting material producing step”, composes the stem-looped DNA that is needed for the second stage. The forward and backward inward and outer primers bind to the complementary sequences within the target DNA sequence[1]. The stem-looped DNA undergoes additional annealing within the target sequences and with the Bst polymerase, which produces stem-loop DNA which concludes the first step[1]. The stem-loop DNA enters into step two- “cycling amplification step”, which uses the internal primers FIP and BIP in hybridization and synthesis reactions to produce an original and a repaired stem-loop DNA strands[1]. For further amplification, loop primers can be inserted into the reactions[2]. The third step is “elongation and recycling”, where the DNA products are recycled and elongated through a BIP-primed strand displacement reaction[1].

Basic schematic of LAMP reaction from Wong et, al (2017)[4].


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References


1. Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N., & Hase, T. (2000). Loop-mediated isothermal amplification of DNA. Nucleic Acids Research, 28(12), e63–e63. https://doi.org/10.1093/nar/28.12.e63

2. Niessen, L., & Vogel, R. F. (2010). Detection of Fusarium graminearum DNA using a loop-mediated isothermal amplification (LAMP) assay. International Journal of Food Microbiology, 140(2–3), 183–191. https://doi.org/10.1016/j.ijfoodmicro.2010.03.036


4. https://sfamjournals.onlinelibrary.wiley.com/doi/10.1111/jam.13647

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