
Quicklypurifyhighyieldsofhigh-molecular-weightgenomicDNA,totalcellularRNAorTotalNucleicAcid(TNA)withonekit
Fast:PurifyTotalNucleicAcid(TNA),DNAorRNAin30–60minutes
- Safe:Doesnotusehazardousphenol,chloroformorguanidine
- HighPurity:A260/A280ratiosconsistentlybetween1.8and2.0
- HighYields:Improvesyieldsbyavoidingtheuseofcolumnswhichoftenreducenucleicacidyields
- Versatile:PurifyTNA,genomicDNA,totalRNA,FFPERNA,orbothgenomicDNAandtotalRNAfromasample
- TotalRNARecovery:Purifybothlargeandsmall(e.g.,miRNA)RNAforRNA-SeqorqRT-PCR
- Proven:HundredsofcitationsforpurificationofDNAandRNAfromdozensofsampletypesforuseinmanyapplications
TheMasterPure™CompleteDNAandRNAPurificationKitenablesrapid,highyieldpurificationofhigh-molecular-weightgenomicDNA,totalcellularRNAandTotalNucleicAcid(TNA)frommanydifferentsampletypes.
Thekitprocedureusesasafe,gentle,andscalablesalt-precipitationprotocol(Figure1)thateliminatestheneedforhazardouschemicalsandyield-reducingcolumns.Purificationreactionscanbescaleduporscaleddowndependingontheamountofnucleicacidneededandthesizeofthesample.MasterPureKitsforNucleicAcidpurificationsGram-positivebacteria,yeastDNAandbloodsamplesarealsoavailable.
Applications
PurificationofgenomicDNAortotalRNAformanyapplications,including:
- Librarypreparationfornextgenerationsequencing(NGS)ofgenomicDNAandRNA
- DNAmethylationstudiesusingIllumina®Infinium®HumanMethylationBeadChips
- GenomicDNAandCDNAcloning
- qPCRandqRT-PCR
- Micrarrayanalyses(CGH,geneexpressionprofiling,etc)
Figure1.OverviewoftheMasterPure™CompleteKitprotocol. |
Figure2.ComparisonofPCRsensitivityusingtemplatesobtainedusingtheMasterPure™CompleteDNAandRNAPurificationKitversusaspin-columnkit.PCRamplificationwasperformedafterextractionfromserialdilutionsofE.colicellsrangingfrom2×107(lanes1)to200(lanes6).LaneM,DNAMarker;lane7,negativecontrol. |
Figure3.DNA,RNA,andtotalnucleicacid(TNA)purifiedfromdiversecellsourcesusingtheMasterPure™CompleteKit.M=kbladder. |
ebiomall.com






>
>
>
>
>
>
>
>
>
>
>
>
这个与琼脂糖电泳有关。提取的DNA分子量很大,可能在40K到100K左右。也有可能在这个范围。但普通的琼脂糖电泳。在大分子的时候根本分别不出来。如果你有DL15000的MARKER你就知道,象那个一万与一万五和条带差别很近。
1、DNA提取问题:现在核算提取试剂盒大部分都是吸附柱法,相信你应该不会操作错误,如果提取之后进行核酸定量结果很低的话,试着再最后一步溶解核算的时候把水稍微加温(37℃就可以了),溶解时间稍微长一些,然后再离心的时候采用分次离心,比如开始你用200ul溶解的话,你可以分两次用100ul水溶解离心,这样可以提高DNA的产率.
2、电泳问题:如果你核酸定量浓度不低的话就考虑电泳问题,电泳的时候加上1kb的marker,如果marker跑不出来说明你凝胶有问题,一般只要marker跑出来了,DNA浓度又比较高,而没有条带这样的现象很少见,DNA提取比较简单而且相当稳定,本人当时做甲基化提取的DNA有一次拿出来电泳忘了放回冰箱了,结果大夏天的在外面放了一天,心怀忐忑的进行了一次电泳,结果条带依然给力,一点都没有降解.
buffer P1:除去RNA
buffer P2:裂解细胞
buffer P3:沉淀DNA
buffer WA、buffer WB:都是洗涤液(这两个之间有什么区别我也不清楚)
TE:溶解DNA.

