Awọn igbimọ ibaramu | 1, 3, 5, 8. |
Sisẹ | fe ase, afamora ase |
Nọmba awọn ibudo abẹrẹ | 5, 6, 7, 8, 9, 10. |
Ibamu awo orisi | C18 awo, jin kanga awo, sintetiki awo (ibaramu pẹlu julọ sintetiki farahan), microtiter awo |
Modulu | ẹyọkan tabi ipo meji |
Foliteji | 220V |
Atilẹyin ọja | 1 odun |
Aṣa | Ti gba |
1. Gel Electrophoresis Chromatography ìwẹnumọ
Lo denaturing polyacrylamide gel electrophoresis chromatography fun ìwẹnumọ.Aṣoju denaturing ni gbogbogbo 4M formamide tabi urea 7M, ifọkansi ti acrylamide wa laarin 5-15%, ati ipin ti methacrylamide jẹ akọkọ laarin 2-10%.
Lẹhin electrophoresis, ipo ti ẹgbẹ nucleic acid nilo lati pinnu labẹ itanna ti ina ultraviolet, jeli ti o ni ibi-afẹde nucleic acid ti ge kuro, acid nucleic ti fọ ati jo, ati lẹhinna ojutu leaching ti wa ni idojukọ, desalted, iwon ati lyophilized.
2. DMT-Lori, HPLC ìwẹnumọ
Yan ipo DMT-Lori lakoko iṣelọpọ, ọja robi ti jẹ centrifuged o si dojukọ ni iwọn otutu yara lati yọkuro amonia pupọ lẹhin aminolysis.
Iyapa ti a ṣe pẹlu lilo iwe C18 pẹlu acetonitrile ati 10% triethylamine-acetic acid (TEAA) bi eluent.Lẹhin ti elution ti pari, o wa ni idojukọ, lẹhinna a ti yọ ẹgbẹ DMT kuro pẹlu trifluoroacetic acid.Lẹhin imukuro, diẹ ninu awọn iyọ ati awọn ohun elo kekere ni a yọ kuro nipasẹ tube ti a ge kuro, ati nikẹhin desalted.
Ọna yii le gba ọja pẹlu mimọ ti o ga julọ, ṣugbọn o jẹ dandan lati fiyesi si iṣẹlẹ ti depurination.
3. DMT-Pa, HPLC ìwẹnumọ
Yan DMT-Paa lakoko iṣelọpọ, ati pe ọja robi naa jẹ centrifuged ati pe o ni idojukọ ni iwọn otutu yara lati yọkuro amonia pupọ lẹhin ammonolysis.
Iyapa ti gbe jade nipa lilo iwe C18 pẹlu acetonitrile ati 10% triethylamine-acetic acid ninu omi bi eluent.Lẹhin ti iyapa naa ti pari ati iwọn, awọn aliquots ti wa ni lyophilized.
Ọna yii nilo atunṣe iṣọra ti awọn ipo iyapa, ati pe o tun le gba awọn ohun elo ibi-afẹde mimọ.