Chizindikiro Chatsopano cha Kutulutsa DNA ya Madzi a Zachilengedwe — Kutsata kwa Bigefei Kumafulumizitsa Kafukufuku wa Sayansi

Kiti iyi imachokera ku dongosolo lokonzedwa bwino la buffer pamodzi ndi mikanda ya nanomagnetic yogwira ntchito bwino kwambiri. DNA ya genomic imagwirizana makamaka ndi magulu ogwira ntchito pamwamba pa mikanda ndipo imalekanitsidwa pansi pa mphamvu yakunja ya maginito. Pambuyo potsuka pang'ono pang'ono kuti muchotse mapuloteni, mchere, ndi zonyansa zina, DNA ya genomic yoyera kwambiri pamapeto pake imathetsedwa.

Chopangidwa mwapadera kuti chigwiritsidwe ntchito poyesa madzi ozungulira, chidachi chimachotsa bwino DNA ya bakiteriya yomwe yasonkhanitsidwa pa nembanemba zosefera, kuphatikizapo mabakiteriya a Gram-negative ndi Gram-positive (mpaka maselo 2 × 10⁹ a bakiteriya pa nembanemba imodzi yosefera). Chimagwirizana ndi makina otulutsa nucleic acid okha kuti agwiritsidwe ntchito kwambiri. DNA yotulutsidwayo ndi yapamwamba kwambiri ndipo ingagwiritsidwe ntchito mwachindunji pa PCR/qPCR, NGS, ndi ntchito zina zotsatizana.

Zinthu Zamalonda

1. Kuthekera kwa Kutulutsa Mabakiteriya Osiyanasiyana

Amachotsa bwino mabakiteriya a Gram-negative ndi Gram-positive m'madzi, kuphimba madera a tizilombo toyambitsa matenda omwe amapezeka m'madzi abwino komanso m'nyanja, ndikukwaniritsa zofunikira zosiyanasiyana zowunikira.

2. Kuyera Kwambiri ndi Kupereka Zokolola Zambiri

Amapereka DNA yoyera kwambiri, yopanda zinthu zoletsa kuipitsidwa, komanso yogwira ntchito bwino ngati mamolekyulu.

3. Kugwirizana Kokha komanso Kogwira Ntchito Kwambiri

Imagwirizana kwathunthu ndi machitidwe ochotsera nucleic acid a Bigfish okha, kuthandizira kukonza zitsanzo 32 kapena 96 nthawi imodzi, kuchepetsa kwambiri nthawi yokonza ndikuwonjezera magwiridwe antchito a labotale.

4. Ntchito Yotetezeka Komanso Yosavuta Kugwiritsa Ntchito

Palibe chifukwa chogwiritsa ntchito zinthu zoopsa zachilengedwe monga phenol kapena chloroform, zomwe zimachepetsa chiopsezo cha chitetezo cha labotale. Zinthu zazikulu zimayikidwa kale m'mabotolo 96, zomwe zimachepetsa zolakwika zogwiritsa ntchito mapaipi ndi manja komanso zimapangitsa kuti ntchito ziyende bwino.

Zida Zogwirizana

Nsomba Yaikulu BFEX-16E

BFEX-32

BFEX-32E

BFEX-96E

Zotsatira Zoyesera

Chitsanzo cha madzi a mumtsinje cha 600 mL chinasefedwa kudzera mu nembanemba, ndipo DNA inachotsedwa pogwiritsa ntchito chida choyeretsera ndi kuyeretsa madzi cha Bigfish magnetic bead chomwe chimagwiritsidwa ntchito poyeretsera ndi kuyeretsa madzi pamodzi ndi chida chogwirizana nacho. DNA yochotsedwayo inafufuzidwa pambuyo pake ndi agarose gel electrophoresis.