Bst 2.0 DNA Polymerase (Glycerol kyauta)
Bst DNA polymerase V2 an samo shi ne daga Bacillus stearothermophilus DNA Polymerase I, wanda ke da 5'→3' Ayyukan polymerase na DNA da kuma aikin maye gurbin sarkar mai ƙarfi, amma babu 5'→3' ayyukan exonuclease.Bst DNA Polymerase V2 ya dace da matsuguni-matsala, haɓakar haɓakar isothermal LAMP (Maɗaukakin madaidaicin isothermal ƙarawa) da saurin jeri.
Abubuwan da aka gyara
| Bangaren | Saukewa: HC5005A-01 | Saukewa: HC5005A-02 | Saukewa: HC5005A-03 |
| BstDNApolymerase V2 (Ba shi da Glycerol) (8U/μL) | 0.2 ml | 1 ml | ml 10 |
| 10 × HC Bst V2 Buffer | 1.5 ml | 2 × 1.5 ml | 3 × 10 ml |
| MgSO4(100mM) | 1.5 ml | 2 × 1.5 ml | 2 × 10 ml |
Aikace-aikace
1.LAMP isothermal amplification
2.DNA strand guda gudun hijira dauki
3.High GC gene sequencing
4.DNA jerin matakan nanogram.
Yanayin Ajiya
Jirgin da ke ƙarƙashin 0 ° C kuma a adana shi a -25 ° C ~ -15 ° C.
Ma'anar Naúrar
An bayyana raka'a ɗaya azaman adadin enzyme wanda ya haɗa 25 nmol na dNTP cikin kayan da ba za a iya narkewa ba a cikin mintuna 30 a 65°C.
Kula da inganci
1.Gwajin Tsaftar Protein (SDS-PAGE):Tsaftar Bst DNA polymerase V2 an ƙaddara ≥99% ta binciken SDS-PAGE ta amfani da gano Coomassie Blue.
2.Ayyukan Exonuclease:Shigar da amsawar 50 μL mai ɗauke da ƙaramar 8 U na Bst DNA polymerase V2 tare da 1 μg λ -Hind Ⅲ narke DNA na sa'o'i 16 a 37 ℃ yana haifar da rashin lalacewa da za a iya ganowa kamar yadda aka ƙaddara.
3.Ayyukan Nickase:Haɓaka amsawar 50 μL mai ɗauke da ƙaramar 8 U na Bst DNA polymerase V2 tare da 1 μg pBR322 DNA na sa'o'i 16 a 37°C yana haifar da rashin lahani da za a iya ganowa kamar yadda aka ƙaddara.
4.Ayyukan RNase:Haɓakar amsawar 50 μL mai ɗauke da ƙaramar 8 U na Bst DNA polymerase V2 tare da 1.6 μg MS2 RNA na tsawon awanni 16 a 37°C yana haifar da rashin lahani da za a iya ganowa kamar yadda aka ƙaddara.
5.E. coli DNA:120 U na Bst DNA polymerase V2 an duba shi don kasancewar E. coli genomic DNA ta amfani da TaqMan qPCR tare da abubuwan da aka tsara musamman don E. coli 16S rRNA locus.Kwafin halittar E. coli na DNA shine ≤1 Kwafi.
Ra'ayin LAMP
| Abubuwan da aka gyara | 25μL |
| 10 × HC Bst V2 Buffer | 2.5 ml |
| MgSO4 (100mM) | 1.5 μl |
| dNTPs (10mM kowane) | 3.5 ml |
| SYTO™ 16 Green (25×)a | 1.0 μl |
| Haɗin farkob | 6 ml |
| Bst DNA Polymerase V2 (Babu Glycerol) (8 U/ul) | 1 μl |
| Samfura | × μL |
| ddH₂O | Har zuwa 25 μl |
Bayanan kula:
1) a.SYTOTM 16 Green (25×): Dangane da buƙatun gwaji, ana iya amfani da sauran rina a madadin;
2) b.Haɗin farko: samu ta hanyar haɗa 20 µ M FIP, 20 µ M BIP, 2.5 µ M F3, 2.5 µ M B3, 5 µ M LF, 5 µ M LB da sauran kundin.
Martani da Hali
1 × HC Bst V2 Buffer, yawan zafin jiki yana tsakanin 60°C da 65°C.
Rashin kunna zafi
80 ° C, 20 min
