Poly A Carrier RNA

Cat Nama: HC4001A

Ko te Poly A, te polyadenylate, he ranunga o te 100 ~ 10000 polyadenylates, he polymerized e polynucleotide phosphorylase in vitro.I roto i te vivo, ka taapirihia te poly (a) ki te 3-terminal o te mRNA e te enzyme hei whakapai ake i te pumau o te mRNA.I roto i te whakamahinga o te tangohanga waikawa karihi, ko te taapiri i te poly A ki te lysate, ki te otinga here ranei ka taea te whakapai ake i te hua o te DNA me te RNA.Ko te tikangao te poly A hei whakapai ake i te hua o te waikawa nucleic e whai ake nei:

1. Whakapaa ki te mata o nga tuhinga.Ko te nuinga o nga tuhinga polypropylene he hiko pateko kei runga i te mata, ka uru ki te waikawa nucleic.Ka taea e te kawe RNA te whakakii i eneite whakangao me te whakaiti i te ngaronga o nga waikawa karihi e whaaia ana.

2. Whakahohe i nga nucleases: He maha nga momo karihi kei roto i nga tauira koiora me tetaiao.Ka taea e te Poly A te whakakore i nga nucleases i roto i nga mahi tangohanga, tiaki ranei ki tete whakapai ake i te tukunga me te pumau o nga waikawa nucleic target.

3. Coprecipitation: I roto i te taahitanga purenga waikawa nucleic o te waipiro takawaenga rerenga, herea ranei, ka taea e te poly A te uru tahi me te waikawa nucleic target, te hanga matūriki polymer raneiwhakapai ake i te whakaora.

Tikanga whakamahi

Tangohia te nui o te paura lyophilized, tāpirihia te wai rongoa DEPC, te wairewa tote guanidine raneimemeha ki roto ki te 0.1-1ug/uL, katahi ka whakaemihia, ka penapena ki te -20°C.

Nga tono

1.Virus DNA / RNA tangohanga: ko te taapiri i te 1-5ug Carrier RNA ki te lysate ka taea te whakapai ake i te hua o te RNA / DNA, te whakapumau i te waikawa nucleic whaainga me te karo i te paheketanga o te waikawa nucleic purea i te wa e rokiroki ana.

2. I roto i te tangohanga DNA/RNA moroiti ma te tikanga membrane pou (<1ug), ko te taapiri i te RNA kawe ki te 1-5ug he pai ki te whakapai ake i te hua o te waikawa karihi.

3. I roto i te waipiro takawaenga waikawa nucleic precipitation me te taahiraa kukū, te tāpiritanga o 1-2ug kawe RNA he āwhina ki te whakapai ake i te whakaora o RNA wāhanga poto.

4. I roto i te otinga tauhohenga PCR tirotiro ine, ko te taapiri i te 10-100ng kawe RNA ki te otinga tauhohenga he awhina hei whakapai ake i te tairongo me te whakaiti i te C_Tuara.