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SFϿ桦ʤ޷Ǽ­Ƥޤ

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ʤ᡼륢ɥ쥹ϡ֤ΤΤyj8.so-net.ne.jpפǤ޻DZѿȾѾʸѴƤɤ


ǽ2006/01/14 03:00

06/01/13:å

֥ͦͦ楦ǥǥǥǥǥLet's Go
4ȢTaqʸ
Ĥ̾PCR
֥ͦͦ楦ǥǥǥǥǥLet's Go
1mlѱդޤ
нκˤϥ
֥ͦͦ楦ǥǥǥǥǥLet's Go
¸Τ˥ޥ򻦤
ȴȴӤĶ
֥ͦͦ楦ǥǥǥǥǥLet's Go
祦祦Х˻
ϥɤ
֥ͦͦ楦ǥǥǥǥǥLet's Go
IJݤ˻
ݤϰɤ
֥ͦͦ楦ǥǥǥǥǥLet's Go
ǥϥ
äƾʪ줤

äȻפĤǡȴطԤˤȽʤͥ⤤ĤפĤޤ

֥ͦͦ楦ǥǥǥǥǥLet's Go
PCR̤ᥢ
ȤbufferMg
֥ͦͦ楦ǥǥǥǥǥLet's Go
PCR󤼤ʤ
äȤĤHotStartͤ


4ȢTaqʸĤ̾PCR
TaqPCRDNAϢȿˤȤDNAפȿιǡȢ001000ʬ١äƤޤĤޤꤳοͤ50ȿʰʾˤPCR򤷤Ƥ׻ˤʤΤǡĤȿ̾ǸƤФ褦ˤʤäȤ̣ܥͥϥꥨ󥿥饸Υ˥ޥԥTVCMΥͥ

1mlѱդޤ롿нκˤϥ
ѱդνϡɤΤ褦ʪޤޤƤ뤫ˤäưۤʤޤúǽꡢºޤȿƤΤƤͤФʤʤѱդ¿ꡢĤ̤ݤȻפäƥӥƤ뤦ˡ´ȤžǸ漼뺢ˤϥӰդѱդޤäƤ롢Ȥ̣ܥͥϡ֥Х䤺餹ץͥ

¸Τ˥ޥ򻦤ȴȴӤĶ
ưʪˡˤäơŪưʪΤιĤˤäˡ¸Ѥưʪɬ׺Ǿ¤򡢶ˤʤ®䤫˻˻餷뤳Ȥ׵ᤵƤޤޥξ硢Ƭˤꤷơĥȡ֤ˤƷǤæޤȱǤΤǡܥޥˤ϶ˤʤзȼʤΤˡշϤݻޤޡˡ®䤫˻˻餷뤳ȤޤηæܿȴӤ¿Ȥáդܥͥޤ

祦祦Х˻Сϥɤ
̤DNAۤϡgΥ祦祦Х򻦤ޤPCRˡȯޤTaqäϤ褯äƤˡ祦祦ХĹ23mmν1mg٤ǤΤǡ1g٤1000ɤȤˤʤޤ̾50ɤ̤ФΤǤޤΥѥȤͥ褷ޤ

IJݤ˻Сݤϰɤ
IJݤDNAФǤϡݤܱաʿml١ˤ
礬¿Ǥʥץ饹ߥɤʤ餽ǽʬ̼ˡξ硢IJݤ1m5mľ췿ʤΤǡѤ10-9mm3̤1mm3mlȤƤ1ɤml100ץФаˤʤʤȤζݤ򻦤Ȥˤʤޤ

ǥϥäƾʪ줤
祦祦ХҤ̤Ĺ23mmΥϥβ˶̼θǽΤ褤롼ڤӤĤ褦ʤΡ˲ǹԤޤ򤺤äƤȡܤʪưʥ㡼䥹饤ɥ饹ưĤĺȤΤǡˤˤؤ餺ΤưʤΤǡ줫λɷȻȾɡλɷ㤬߹鷺ʪ줤ˤʤġĤΤϻ

PCR̤ᥢȤbufferMg
PCRȿդˤMg2+󤬴ޤޤƤޤ̾1.52.0mMˡźդbufferʴ˾ױաˤˤMg2+󤬴ޤޤƤΤȴޤޤƤʤ(Mg FreeˤޤǤȿ˴ؤMg2+Ϳƶ礭ΤǡȿѤȤˤMg Free bufferMgCl2ϱդʬǺĴޤְ㤨MgbufferˤMgCl2ϱդäƤޤMg2+ǻ٤⤯ʤꤹðŪDNA褦ˤʤޤΤᤵޤޤĹDNAҤƤޤŵưdzǧȡðŪʥХɤΤǤϤʤ̵ΥХɤ졼ΤܤäޤäƸ֡ʥᥢˤˤʤäƤޤȤá

PCR󤼤ʤäȤĤHotStartͤ
PCRȿѤDNAǤǮΤΤǤϤ뤬ȤϤ٤95DNAܺ򤤤ưܺˤdenatureλβ١30á1ʬ2040١ˤ˲ǮȤȳ򼺤äƤ椯ΤǤ롣̾PCRȿDenature³5070anealingȸƤФprimerDNAðŪΰؤ她ƥåס72DNAȿextention˥ƥåפΡ3ĤΥƥåפ򷫤֤ԤðŪΰΤߤܤ䤷Ƥ椯ΤǤ롣ǽΤۤη֤ðŪΰ褬ƤޤȤܤƤޤΤǡðŪΰΤߤ䤹ˤϺǽ餬פǤ롣
ϱդĴɹ4ˤǹԤǽdenature95ˤޤǾ夬֤anealingextentionβ٤̲᤹뤳Ȥˤʤꡢ㤤١ðŪΰؤprimer夬뤳Ȥ⤢롣ɤᡢDNAǤι¤򾯤Ѥ9510ʬʤɤιǮǤƤϤƳĤ褦ʷˤǤΤƤꡢHotStartȸƤФƤ롣ΥפιǤȿ򤹤Ȥˤϡƥåפη֤9510ʬʤɤβǮԤץȤΤ̾ιǤʤΤ˴ְ㤨ƤΥץȤäƤޤȡ10ʬβǮδ֤˹Ǥ褷Ϥᡢȿ٤֤ǤȿˤʤäƤޤPCRʪ̤äƤޤȤԤäȤá

ϥեǤꡢºߤοʪȤϲط⤢ޤ



06/01/12:ǽξ

primaly cell culturegooŵľƤߤޤˤ϶ܤϤꤽǤʤȤʤˡ



06/01/11:δ

ǯνؤǵĤȡ
Ϻǯä
ǯǤ롣

Ĥޤꡢ30ξ񢪸ʿǯ񢪾񢪸פ³㤦餷Τ Τˤǯ塢лŪˤϤۥ󤬶ꤹ㹹ǯ㳲ΤʿѼ̿ξƤ˴ʤʬ³ǯǤϤޤɤ͡



06/01/10:ޤܲ

Ͼ֤äŹ1ְʾ줿ˤΤǡ㤤ƨƤܤҤġŹĹ󤫤ŹΥɥ󥯥åȤäƤޤޤ̡ϡɥСʤäΤˡʤʶۤˤʤΤ𡨥Ʋ1߰ʾ㤦ȥӥåȤ򤯤
114ڡˤˤϻΥȡå󤬤뤽ǡäȶ̣ġ

ȤСۥΥ祦祦ХǤ롣
ϥ祦祦ХD. melanogasterˤΤޤޤʹưѰۤ򸡺ƤơФᰦưʤͺ򸫤ĤΤǤ祦祦ХˤưϡޤͺĤĵᰦΡLove songˤͭ¤οưդǤơʹ󤬤äȤꤷߤ٤ʤäȤǡͺӤʥåԥ󥰡ˡ󤬻ߤޤäȤǸ˻ȤήˤʤäƤޤʤʤߤϵᰦΤİƤΩߤޤ餺ͺݤ⤭ˡΤᡢͺꡢäƵᰦΤդǤʤȡҶʤȤˤϽޤ
¿ѰΤͺȻɤŤĻɤˤ졢ưΰʾ夬ʤåƤޤ줤Ǻޤ줿ɤΤޤ󤬡ĤˤФ̣򼨤ʤͺ򸫤ĤΤǤ
äȤηη򳫤ȤؤǺΤƵäƤ˰㤤ʤȻפäϡѰ۷satori̾դΤǤ
̾祦祦Хηϡ1ܤӤ˿ɤʾͺȻƻޤˤƤ⡢¡äʪȤȯڤʪȤݤäפΥѥȤ磻Ȥ繥Ǥ㥷祦祦Хˤμˤʣͺ󤬤Τ̤ǤΡ̡פ˶ᤤʣͺƱ路ƤĶˤ줿satoriѰ۷ͺϡͺФƵᰦư򼨤ΤǤ
ġsatoriϸäƤΤǤϤʤñʤۥ⡢ȤĤƤޤäΤǤ

äǤ͡ηΥ祦祦Х˴ؤƤۥ֤֡Ӥdzڤʤۥ⥹ˤȤäƤϤޤäǤġ̵ȤϤ狼äƤƤ˥ҥȤΥۥʤư˶ޤ͡

ʤߤˤsatoriθҤfruitlessƱꤵƤޤfruitlessZnե󥬡¤DNA祿ѥ򥳡ɤƤꡢΰҤѰۤȡѰۤμˤäfruitlessȤͺͺξư򤹤ξȤХΥϥǤ͡Ѱ۰Ҥ䡢satoriΤ褦ʥۥΥϥˤʤäƤޤȤȤ餫ˤʤäƤޤ
ʾܤȡFruѥǾ˶ɺߤк˦κ˦򥳥ȥ뤹뤳ȤǾͺðŪʿвϩ˴ؤ롣祦祦Хξ硢ΡXˤȾΤΨΰ㤤ͺˤȯѥΰ㤤Ƴ졢ɾήΥѥƯfruitlessΥץ饤󥰥ꥢȤޤ졢FruѥƯΰ㤤˴ؤȤƤ

¾ˤǤspinsterˤʤƤѰۤ⸦椷ƤʤΤǡäʹΤǤϤʤȡ



06/01/09:Τ˿겾̾򿶤뤽ο

־ǦˡġץåȡҾ֤󤫼Ҥ饳ߥåʸˤ줿쥳ߥåΡ־ǦˡġפȡФιפƤƤǤʰ ϤΡФιפκǸ˼夯ơޤӤ륷ʤΤ˵㤱־Ǧˡġפ⤹񤭤ǡ֤ʤס֤ȤפϿƤơѤʤȻפˡ ġĤĴҤǡֽν͡פȤΥˡפȤʤʡǤäƥå꡼³ФꤹȤ줷ʡȵäƤߤ롣 .comΥȤ



06/01/08:Ĵ

β岼ʤʤäƤʴ뤵ʤŻ̮郎ʬ120ĶʤˡȤϤɤΤϤ꤬ʤΤǽȤǤƲᤴ
ȯǮΤȽäƤϤƤ⡢Τ᡹ˤΤϤäǯäݤäƤʡ



06/01/07:Ȥ餷

ǮѤ餺ξ岼ư򷫤֤򤤤餤ŪǮ夬äƲΤǤ⤦Ū;͵ä饰դʤɤĤ餤⥤ɥ֤Ǿ徺ΤŪ˾徺ΤȤ徺ȲߤоŪʤΤɤȤȤ

ޤǤβפäƤץڥ塼ϥӥˤ㤨Ƥ⡢βפϥåȤǤʤʤ



06/01/06:µΤȤϤäȿ

ָʤȤϤȿµΤȤϤäȿפȤΤȡ*1ˤβȷäȵƤޤɡФƤΤǤǤˡ
⤽¹ԤƤޤߤʤʤäƤߤ¤Ͽ٤롣¤ܤΤ򡢤äȡ롣ʤäꡢǤäꤹ褦ʤ鿲ưʤ
Ȥ桼ץǽưǾڤǤȯǮ˿äƲǮޤޤͤФȻפä֤٤äȤΤ⤢Ǥġľ*2ˤäƤΤϽŤ(^^;)

*1֥ХӥޤDzޥ롩פΥҥǡ֥ѥפ򿩤٤褦ȤȤ졢Ȧ
*2鷺Τ줿ᤷʺڤᤷᥤˤΥŹƼ꿩䥫졼ʤɡ˥塼Ū˭٤ǡ24ֱĶȡΥ祤ڽ꿩ήФˤӤϻĤޤˡ



06/01/05:ǥꥺ

2롣ҤɤơĤƤޤ٥åɤξۤĤޤäƿ̤롣
ζǡʸ6ɤƤ4Ⱦˡưŷնˤ㤯ޤϤδ֤ʤɽ뤯ۤ򽳤Ф餤Ƥ򡢺Ƕˤơʤϼưˡ28ʤ25١ˤˤƤ⡢̤Ȥޤʤơե꡼夳ǡޤФ٤ɤ䤷ĤĿ̤Ƥ
1֤餤ǡ褦䤯ΤޤäΤ̤ߤޤäơä̲롣

ġĽ뤵ܳФ롣6
ڤƵפʥޡ1֤ǻųݤˤ̵ʤȤο᤯ˤǡλ֤˽뤤ϤϤʤæե꡼æۤФƤʤ뤤䶻٤äȤǨƤ롣

餫ˤ

Ĥޤϡ2ȯǮ֤ݤäƤβ6˲Ϥ᤿褦ΤΤ᡹̯ˤäż֤ǥӡ뤬ʤޤʤä850mlޤʤäΤˡġĤǼ

Ϥ뤬äĤϤ߽ФƤ֤ǿΤäƤȤǡǤ⿲ʤϤʤäƤȤǡġʾ֡ۤޤä٤ˤäΤ78줫ä̲뤳Ȥ褿ΤǤ
ۤƱȯǮ¬ġijꥺ϶äƤʤȤǧ줿ΤǤ
ȼ򤸤ʤǮްǤ鿲ȤΤˡ



06/01/04:Ĥʹ֤ˤʤޤ

нءŷäضȤǡʼˤޤ褦ˤȤδꤤƤΤȤ
ǤäƥХˤäƹԤäΤġ١äåäΤǡƻäƤäΤä

դȥ硼ɥ˱Ǥ뼫ʬ򸫤ơʹ֡ס
ٺ𱩤ӽФƤΤǡޤ˿夫鿴¡ȴ줿ʤʹ־֡ġĤäƳϿ¡ϼȴƤޤǤɤ͡

ХˤĤȤˤ⤵󤶾ФΤˤʤꡢġĤä2ƻԤͤ˽Фץ쥼ȤΤȤǼ롣



06/01/03:Х

Х档ʤᤢޤĶȤƤŹʤƤӤ
ۥƥ餷ϲŬȤ뤷ĥäƤäǤ뤷٥åɤϼ­򿭤Ф̲뤷˳ڶ˳ڡ



06/01/02:줫β

줫᡼뤬Ϥǯΰ³ơֺǯϤޤʤ褦ˤ͡פȤä
ġĤǤˤϼˤޥޥ



06/01/01:ǯϽ

ԤĤ20ʬ٤ŷؤꤹ롣Х˹ʵꥰå򤤤ĤΤޤ޿ˤޤꤷơITͤį롣
ǡβǰաġ̣⤤餻(^^;)
⤦ա⤦äȰ⤦ʤɤȸĤĤɤ夤ΤϽոΥѥήФ˰դʤä᤯Υ饪ۤˤ⤰ꤳळȤ饫饪

Ǥ2֤ǤơȽ뱿žż֤DzȤ˵äȡ





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