Construction of long-transcript enriched cDNA libraries from submicrogram amounts of total RNAs by a universal PCR amplification method.
[No authors listed]
摘要
Here we report a novel design of linker primer that allows one to differentially amplify long tracts (average 3.0 kb with size ranges of 1-7 kb) or short DNAs (average 1.5 kb with size ranges of 0.5-3 kb) from a complex mixture. The method allows one to generate cDNA libraries enriched for long transcripts without size selection of insert DNAs. One representative library from newborn kidney includes 70% of clones bearing ATG start codons. A comparable library has been generated from 20 mouse blastocysts, containing only approximately 40 ng of total RNA. This universal PCR amplification scheme can provide a route to isolate very large cDNAs, even if they are expressed at very low levels.
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基因功能
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基因
Oog3, Gm12000, Tmem254c, Gm2381, Gm13306, Gm2506, Ccl27b, Gm3286, Gm13242, Rps15a-ps6, Rex2, Zfp955b, Zfp850, Camta1, Srp54c, Uckl1os, AU019176, Slc44a1, 9030411K21Rik, E330021D16Rik, 2810013P06Rik, Zfp534, 2900076A07Rik, Gm15008, AA792892, Tyw1, Nfxl1, Ttll3, Gm29811, Grwd1, Spty2d1, Phkb, Arhgef18, Cog4, Gm15241, AU023762, Bbs4, AU022751, Pan2, Cep57l1, Mbtd1, Psme4, Tmem98, AI503301, Wdr92, Synj1, Itga9, Adcy3, Dgkz, Tsr1, Ptpn23, Epdr1, Golm1, AA986715, Prkaa1, Nckap1l, Espl1, Osbpl11, AI317158, Ankrd12, Hmgxb3, Gramd3, Me2, Eprs, Lgr4, Wwp1, Scamp1, Pacs1, Shmt2, Camk2d, Cstf2, B3gnt5, Galnt7, Mthfd1, Fam210a, Ankhd1, Nusap1, E2f8, Tpr, Igip, Orai1, Cdcp1, Braf, Tmbim6, Shroom2, Prdm2, Rps6ka3, Srsf1, Lims1, Cds2, Stmn1-rs1, 2610305D13Rik, Abcb7, Slc33a1, Npnt, Macf1, Rpl31, Rasa2, Spred1, Acvr2a, Psg29, Ddhd1, Dcun1d1, Adh5, Aebp2, Aga, Agrn, Aim1, Alox12e, Mta1, Tgs1, Ank3, Ap1g1, Ap3b1, Ap4s1, Apc, Arhgap5, Atf2, Atp5a1, Bcl2l1, Bmp8b, Bmpr1a, Bop1, Hyou1, Cacnb2, Pdia4, Caml, Capg, Cbx5, Ccnd2, Cd2ap, Entpd6, Cd82, Cdc42, Cdk6, Cebpg, Inadl, Clcnka, Cnbp, Col18a1, Col4a5, Cops2, Cpt2, Crmp1, Csnk2a2, Vcan, Celf1, Cyp17a1, Cyp27b1, Cd55, Dag1, Dbnl, Dclk1, Dctn1, Dcx, Asap1, Diaph1, Dlg1, Dmwd, Dnm2, Dyrk1a, Mapre1, Rhbdf1, Eif4a2, Eif4g2, Enc1, Ankrd6, Ubox5, Sec63, Bmp2k, P2ry14, Ttbk2, Srsf10, Fbrs, Fech, Fem1b, Fmn1, Fn1, Fnbp1, Srgap2, Fxr1, Galnt1, Gata6, Mtpn, Gcnt2, Gfpt2, Gli1, Gna11, Gnai3, Rack1, Lrp2, Gpc4, Trip12, Hdac6, Mfsd14a, Hipk2, Hnrnpk, Hsd3b4, Hsph1, Ica1, Igf2r, Itgb1, Itpr1, Itsn1, Jup, Kcnh1, Kcnj3, Kif11, Kif13a, Kif13b, Kif2a, Kifc2, Kin, Klf12, Klf4, Kpna3, Krt84, Arhgef1, Ldlr, Lhb, Sertad3, Ripply3, Smad2, Havcr1, Il17rd, Mfap2, Matr3, Meg3, Men1, Mest, Aff1, Mre11a, Msh5, Map4, Map6, Usp34, Ppp1r12a, Myt1l, Ncoa2, Neurod1, Nfe2l1, Nfyc, Nmt1, Zfp638, Npc1, Nsd1, Sqstm1, Mybbp1a, P2rx7, P4ha2, P4hb, Pcnt, Pdcd6ip, Pde4b, Peg3, Rhox5, Pfkl, Pik3cd, Pik3r1, Pja1, Prkcz, Plaa, Papola, Plcg1, Pld1, Plec, Pa2g4, Pln, Plp2, Pml, Pms2, Pold2, Polg, Ppargc1a, Psap, Psmb6, Flna, Ash1l, Hps1, Ptpn13, Ptprc, Rapgef6, Ptprr, Itgb4, Purb, Slu7, Oog1, Fam65b, Rab3d, Rad17, Gm13023, Dcdc2a, Utp14b, Rbm3, Rbmx, Rbpj, Rlim, Rock1, Rp2, Rps24, Trim30a, Scd1, Scd2, Sdf4, Glg1, Sept8, Sh3gl3, Shmt1, St6gal1, St3gal2, Slc1a3, Slc20a1, Slc2a1, Slc6a2, Smarcc1, Snap23, Sod1, Sall3, Spin1, Sptbn1, Srebf2, Esrp1, Urb1, Btla, Dot1l, Stag2, Sgms1, Stat1, Stat6, Adamts18, Stxbp4, Supt4a, Gen1, Syt1, Lpp, Irgq, Mettl14, Tbc1d31, Mkx, Fam91a1, Sepsecs, Pank3, Tmem18, Hk3, Klhl32, A730008H23Rik, Dse, BC016579, Szrd1, Tapbp, Map9, 9530068E07Rik, Tcea1, Zcchc6, Ubtf, Cdk5rap2, Fam168b, Rhot2, Fam20b, Slc43a2, Rab11fip3, Rap1b, Zmat1, Trappc10, Sbno2, Aldh1l2, Tcp11l2, Mars, Mfap3, Gemin5, Tom1l2, Tec, Chd3, Camta2, Taok1, Gm11545, Kat7, Cdc27, G2e3, Baz1a, Tex2, Tex264, Zfand3, Sipa1l1, Rcor1, Rapgef5, Larp4b, Heatr1, Tgfbr1, Tgif1, Phactr1, Nup153, Kdm1b, Tia1, Fcho2, Timeless, Timp3, Atp6v0a2, Tjp2, Sec24c, Tle4, Wapl, Fermt2, Dlgap5, Mtmr6, Tnfaip3, Tnfrsf9, Dedd, Tpm1, Trh, Ctr9, Ttc3, Ttn, Tub, Ubc, Ube2i, Ubp1, Unc5c, Usf2, Utrn, Farp1, Stk24, Fam105a, Ezr, Vill, Mkl1, Wars, Baz1b, Pi4ka, Golgb1, Dzip3, Cldnd1, Flywch1, Uhrf1bp1, Plcl2, Zfp959, Safb2, Dus3l, Lclat1, Fez2, Fbxo11, Ammecr1l, Pggt1b, Atrx, Xpa, Ercc5, Oosp3, Patl1, Trpm6, Abhd17b, Erlin1, Pprc1, Siae, Atrnl1, Ywhaz, Zranb3, Zbtb41, Suco, Trim26, Cnst, Ahctf1, Mark1, Zscan21, Slc39a10, Zfp51, Tmem194b, Zfp52, Zfp59, Zcchc2, Zfp97, Man1b1, Qsox2, Sec16a, D2Wsu81e, Nup188, Sestd1, Madd, 1110051M20Rik, Gyltl1b, Zfp341, Slc17a9, Arfgap1, Zbtb10, Ythdf3, Acad9, 4932438A13Rik, D930015E06Rik, Tmeff1, Prpf38a, Foxj3, Zmpste24, Zbtb40, Tardbp, Kmt2c, Polr2b, Grsf1, Cox18, Cnot6l, Ccz1, Lmtk2, 4933427D06Rik, Ppp4r2, Zfp637, Wnk1, Zfp212, Vps33b, Ppfia1, Ddx49, Colgalt1, Cnot1, Dync1li2, Spg7, Zfp809, Nfrkb, Herc1, Plxnb1, Zfp445, Pyhin1, Stard8, Gas2l3, Gpatch8, Akt3, Apob, Ash2l, Cog5, Akap6, Def8, Dido1, Dmtf1, Ets1, Farsb, Tnpo1, Ogdhl, Fam160b2, Pacsin2, Klhl6, Arid1b, Tiam2, Cchcr1, Ik, Abce1, Ccdc85b, Slc35a1, 9930021J03Rik, Srp54a, Teddm1a, Tor1aip2, Tlk2, Pgap1, Zbtb6, Zfp68, Fam171b, Phc3, Gm4961, A430033K04Rik, Kbtbd8, Parp12, Zfp568, Chd2, Npat, Peak1, Nlgn3, Ift52, Oas1h, Cd300lf, Trim61, Adgre5, Map2k4, Map3k3, Mapk1, Orc4, Ncdn, Irak4, Zbtb24, Sh3pxd2b, Zkscan17, Zfp652, D430019H16Rik, Cln8, Cops7a, Ddx3y, Cntrl, Setx, Ppp2r5c, Ppp2r5e, Vps39, Jade1, Serinc3, Rccd1, AU019823, Ncoa4, Eif2ak4, Tstd2, Trp53bp1, Csnk1e, Abcf3, Mkln1, Amot, Tango2, Fam21, D7Wsu130e, Cep63, Flnb, Mga, Smtn, Ulk2, Mlf2, 9330112F22Rik, D330005C11Rik, Zfp750, Zfp182, Fam168a, Zfp944, Nt5dc1, Eml5, Cep295, Ablim3, A130004G11Rik, E030042N06Rik, Mpzl3, Sbf2, A630001G21Rik, Uggt1, Rps6kc1, Zdhhc17, Fubp3, Ubn2, B930068K11Rik, D630008O14Rik, Ifitm10, Dcaf5, Gpr183, Dna2, Lcmt2, Nol4l, Aknad1, Fam102b, Taok3, Zfp82, Tgm4, Thoc2, Zscan10, Mia3, Map3k9, Fa2h, Edc3, Obscn, AA536875, Tmem106c, Hjurp, Accsl, C87499, 2610005L07Rik, N4bp2l2, Pabpn1l, Crtc1, Cep162, Fbxw24, D230040A04Rik, Zfp950, Itprip, Cpsf6, Akr1c19, Zfp708, Sgpp2, Gm13154, Trim34b, Gm5760, Thop1, Ercc4, Ero1l, Tmprss2, Atp11a, Copb2, Exosc10, Myadm, Hnrnpdl, Nasp, Cpsf2, Atg13, Rragd, Atxn1l, Wwc2, D6Ertd527e, Sept11, Tet1, Mettl17, Krr1, Slx4, Ahi1, Psmd10, Tsnax, Lpar2, Spag5, Copg2, Cd160, Epb41l4b, Ctnnal1, Fmn2, Nfat5, Iqgap2, Wdfy4, Olfr288, Baiap3, Abhd2, Gramd2, Tfip11, 2610042L04Rik, Ap3m1, Tmem131, Btg4, Set, Pfpl, Ptbp2, Ddx21, Acin1, Zfp386, Rassf1, Aatf, Net1, Kcnj1, Letm1, Stx5a, Pdcd10, Socs5, Sept6, Ccnl1, Mbnl1, March7, Tmem183a, Ehd3, Tacc2, Eral1, Zfp318, Rad18, Fam184b, Repin1, Rab2a, Huwe1, Stk25, Nsa2, Erbb2ip, Ncstn, Iigp1, Fads3, Actn4, 1700028E10Rik, Gm13051, Gm15801, Kcnq1ot1, Slc29a1, Gng13, Dhx38, Inpp5e, Sdr39u1, Cubn, Knop1, Rsl1d1, Rpl15, Rps15a-ps4, Gm14393, Srp54b, Rpl31-ps12, Bod1l, Esf1, Frmpd1, Vps13b, Spcs2, Sltm, Tmed7, Gapvd1, Gm10416, Gm8615, Gm8668, Zfp600, Gm8801, Smim7, Bin2, Hint3, Zfp507, Mfsd1, Gm9403, Lman2, Trim59, Rpp14, Paics, Polr3d, Cand2, Ttc14, Mastl, Nop56, Mtdh, Zmynd19, Ccdc77, Smc6, 2810474O19Rik, Fam69a, Ccdc53, Cltc, Zc3h13, Herc4, Paxbp1, 4833420G17Rik, Erp29, Dhdds, Adck3, Ccdc127, Ikbip, Snap29, Ccdc50, Ttll4, Lrrc18, Ube2r2, Nob1, 4930523C07Rik, Alkbh8, Pop1, Slc38a2, Arpc5, Ilf2, Rnmt, Fam32a, Mesdc2, Mphosph10, Tmx3, Zwilch, Chac2, Aen, Ino80, Wls, Spire1, Fam135a, Rnaseh2c, Rbm28, Rpa1, Ddx39, Mto1, Rab14, Tlcd1, Lmbrd1, Arel1, Tab2, Trim37, Angel1, Anln, Phtf2, Mypn, Nexn, Ddi2, Foxk2, Rnf122, Chst8, Fam216a, Ctc1, Nol11, Anapc13, 1810013L24Rik, Sec31a, Snx5, 2610030P05Rik, Wdfy1, Zfp932, Dab2ip, Lnp, Ipmk, Fbxo7, 1600015I10Rik, Fyttd1, Vwa9, Trnt1, Ube2q1, Ogfrl1, 2310068C19Rik, Poc1a, Psmd1, Rpf1, Pigw, Gpatch2l, Ints2, Csnk1g3, Inf2, Unc13d, Rprd1b, Cpne3, Tbccd1, Zc3h11a, Pak4, Nolc1, Cep192, Fancd2os, Mier1, Asun, Arhgap26, Ano9, Arid5b, Pex1, Arhgap21, Ankrd40, Bcor, Sfpq, 9030624J02Rik, Afmid, Tmem248, Rnf215, Osbpl3, Klhdc3, Klhl5, Ankle2, Ptcd1, Lancl2, Esco2, Slc35f2, Ddx42, Kdelc1, 2010111I01Rik, Rap2c, Cnot2, Cenpn, Pgm2, Fryl, Amer1, 2310050P20Rik, Prr5l, Hsdl2, 2610202C22Rik, 2610319H10Rik, Rps6kb1, Tagap, Pan3, Zfp655, Dis3, Zfp74, Fam76b, Setd5, 2900006B11Rik, Ccser2, 2900084O13Rik, Pcdh18, Arhgef6, Armt1, Rnf38, 2410089E03Rik, Kif2c, Otud4, Dnm1l, Als2, Glyr1, Nphp3, Nol9, Slc7a13, Armc8, F13a1, Dtx2, Cyld, Hopx, Ubap2l, 4933427D14Rik, Lrrc71, Sost, Lrrc17, Fam234b, Gvin1, Scrn3, Zfp943, Rhoh, C2cd5, Platr14, Fam63a, Uhrf1bp1l, Asxl2, Parpbp, 4930599N23Rik, Fam35a, Eif4b, Dcaf17, Smurf1, Zfp821, Dcp1a, Cant1, Arhgap15, Depdc1a, Erp44, Pcnp, Cep112, Clasp2, Ube2d-ps, Srxn1, Trim12a, Snx27, Klhdc10, Dtl, Spopl, Cers2, Ticrr, Sun1, Tmem181a, C330018D20Rik, Usp45, Zfp955a, Ralgps2, Ddx52, Pym1, Brd8, C330013J21Rik, Rictor, Ltn1, Saal1, Cdk5rap3, Wwox, Nfkbiz, Ckap2, Ankrd17, Akap12, Bicc1, Mmp1b, Aff4, Pard3, Brwd1, Clmn, Spock2, Sfxn3, C87977, Nlrp4f, B230337E12Rik, D2hgdh, 1700034H15Rik, Hnrnpf, Nat10, Qser1, Dnttip2, Ankrd50, Arfip1
原始数据
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文献解读
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