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- 详细信息
- 技术资料
- 保存条件:
-20
- 保质期:
2年
- 英文名:
pRL-TK
- 库存:
100
- 供应商:
上海烜雅生物科技有限公司
- 规格:
干粉/液体
基本信息
名称:pRL-TK哺乳报告质粒
别称: pRL-TK
质粒属性
质粒简介
The pRL-TK Vector may be used in combination with any experimental reporter vector to cotransfect mammalian cells. However, it is important to realize that trans effects between promoters on cotransfected plasmids can potentially affect reporter gene expression. This is primarily of concern when either the control or experimental reporter vector, or both, contain very strong promoter/enhancer elements.The occurrence and magnitude of such effects will depend on several factors: i) the combination and activities of the genetic regulatory elements present on the cotransfected vectors, ii) the relative ratio of experimental vector to control vector introduced into the cells, and iii) the type of cell transfected.
质粒图谱
质粒序列
LOCUS Exported File 4045 bp ds-DNA circular SYN 01-4-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pRL-TK
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4045)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-4-1 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4045
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 8..759
/note="HSV TK promoter"
/note="herpes simplex virus thymidine kinase promoter"
intron 829..961
/note="chimeric intron"
/note="chimera between introns from human?beta-globin and
immunoglobulin heavy chain genes"
promoter 1006..1024
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2000..2121
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2254..2358
/gene="bla"
/note="AmpR promoter"
CDS 2359..3219
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3390..3978
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
61 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
121 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
181 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
241 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
301 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
361 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
421 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
481 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
541 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
601 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
661 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
721 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc ttctgacaca
781 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
841 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
901 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
961 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
1021 taggctagcc accatgactt cgaaagttta tgatccagaa caaaggaaac ggatgataac
1081 tggtccgcag tggtgggcca gatgtaaaca aatgaatgtt cttgattcat ttattaatta
1141 ttatgattca gaaaaacatg cagaaaatgc tgttattttt ttacatggta acgcggcctc
1201 ttcttattta tggcgacatg ttgtgccaca tattgagcca gtagcgcggt gtattatacc
1261 agaccttatt ggtatgggca aatcaggcaa atctggtaat ggttcttata ggttacttga
1321 tcattacaaa tatcttactg catggtttga acttcttaat ttaccaaaga agatcatttt
1381 tgtcggccat gattggggtg cttgtttggc atttcattat agctatgagc atcaagataa
1441 gatcaaagca atagttcacg ctgaaagtgt agtagatgtg attgaatcat gggatgaatg
1501 gcctgatatt gaagaagata ttgcgttgat caaatctgaa gaaggagaaa aaatggtttt
1561 ggagaataac ttcttcgtgg aaaccatgtt gccatcaaaa atcatgagaa agttagaacc
1621 agaagaattt gcagcatatc ttgaaccatt caaagagaaa ggtgaagttc gtcgtccaac
1681 attatcatgg cctcgtgaaa tcccgttagt aaaaggtggt aaacctgacg ttgtacaaat
1741 tgttaggaat tataatgctt atctacgtgc aagtgatgat ttaccaaaaa tgtttattga
1801 atcggaccca ggattctttt ccaatgctat tgttgaaggt gccaagaagt ttcctaatac
1861 tgaatttgtc aaagtaaaag gtcttcattt ttcgcaagaa gatgcacctg atgaaatggg
1921 aaaatatatc aaatcgttcg ttgagcgagt tctcaaaaat gaacaataat tctagagcgg
1981 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
2041 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
2101 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
2161 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
2221 aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
2281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
2341 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
2401 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
2461 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
2521 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
2581 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
2641 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
2701 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
2761 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
2821 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
2881 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
2941 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
3001 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
3061 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
3121 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
3181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
3241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
3301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
3361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
3421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
3481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
3541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
3601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
3661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
3721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
3781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
3841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
3901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
3961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
4021 tggccttttg ctcacatggc tcgac
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后,请先根据产品管壁标签来判断产品形式,并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉(常温运输,存于-20度,90天保质期,请务必转化挑单克隆培养,不要直接使用和测序)
①收到质粒干粉后请先5000rpm离心1min,再加入20μl ddH2O去离子水溶解质粒;
②取1支100μl 感受态于冰上解冻10min,加入2μl质粒,再冰浴30min后,42℃热激60s,不要搅动,再冰浴2min;(从第二步开始均要在超净工作台中无菌操作)
③加入900μl无抗的LB液体培养基,180rpm震荡37℃培养45min;
④6000rpm离心5min,仅留100μl上清液重悬细菌沉淀,并涂布至目标质粒抗性的LB平板上;(可使用本平台的平板涂布专用玻璃珠进行涂布,可以比传统涂布方法获得更多转化子)
⑤将平板正向培养1h,再倒置37℃培养14h。如果要求是30度则培养20h;
(菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态,要先转克隆感受态,重提质粒后再导入表达感受态)
⑥挑取单菌落至LB液体培养基中,加入对应抗生素,220rpm震荡培养14h,根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种(冰袋运输,存于-80℃,保质期90天,请务必划线挑单克隆培养)
四区划线后挑单菌落培养,酵母菌需要先液体复苏再四区划线,再挑单菌落液体培养。
3、穿刺菌种(冰袋运输,存于4℃,保质期7天)
穿刺接种,液体培养后四区划线,再挑单菌落液体培养。
4、菌落平板(冰袋运输,存于4℃,保质期7天)
直接挑取单菌落至液体培养基中。
5、液体质粒(冰袋运输,存于-20℃,保质期90天)
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒(常温运输,存于-20度,90天保质期,请务必转化挑单克隆培养,不要直接使用和测序)
收到货后将滤纸画圈部分剪下放入EP管中,加100ul无菌水将滤纸浸湿并浸泡5min,吸取5ul质粒转化,离心全涂。
二、转化图片
名称:pRL-TK哺乳报告质粒
别称: pRL-TK
| 启动子: | TK |
|---|---|
| 复制子: | pUC |
| 终止子: | SV40 poly(A) signal |
| 质粒分类: | 哺乳细胞,信号通路报告载体 |
| 质粒大小: | 4045bp |
| 原核抗性: | Amp |
| 克隆菌株: | DH5a |
| 培养条件: | 37度 |
| 表达宿主: | 哺乳细胞 |
| 诱导方式: | 无须诱导,瞬时表达 |
| 5'测序引物: | T7:TAATACGACTCACTATAGGG |
| 3'测序引物: | 根据序列设计引物 |
质粒属性
| 载体宿主: | 哺乳细胞 |
|---|---|
| 载体用途: | 信号报告 |
| 基因种属: | |
| 基因类型: | |
| 原核抗性: | Amp |
| 真核抗性: | |
| 荧光蛋白: | rLuc |
质粒简介
The pRL-TK Vector may be used in combination with any experimental reporter vector to cotransfect mammalian cells. However, it is important to realize that trans effects between promoters on cotransfected plasmids can potentially affect reporter gene expression. This is primarily of concern when either the control or experimental reporter vector, or both, contain very strong promoter/enhancer elements.The occurrence and magnitude of such effects will depend on several factors: i) the combination and activities of the genetic regulatory elements present on the cotransfected vectors, ii) the relative ratio of experimental vector to control vector introduced into the cells, and iii) the type of cell transfected.
质粒图谱
质粒序列
LOCUS Exported File 4045 bp ds-DNA circular SYN 01-4-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pRL-TK
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4045)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-4-1 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4045
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 8..759
/note="HSV TK promoter"
/note="herpes simplex virus thymidine kinase promoter"
intron 829..961
/note="chimeric intron"
/note="chimera between introns from human?beta-globin and
immunoglobulin heavy chain genes"
promoter 1006..1024
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2000..2121
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2254..2358
/gene="bla"
/note="AmpR promoter"
CDS 2359..3219
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3390..3978
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
61 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
121 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
181 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
241 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
301 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
361 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
421 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
481 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
541 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
601 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
661 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
721 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc ttctgacaca
781 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
841 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
901 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
961 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
1021 taggctagcc accatgactt cgaaagttta tgatccagaa caaaggaaac ggatgataac
1081 tggtccgcag tggtgggcca gatgtaaaca aatgaatgtt cttgattcat ttattaatta
1141 ttatgattca gaaaaacatg cagaaaatgc tgttattttt ttacatggta acgcggcctc
1201 ttcttattta tggcgacatg ttgtgccaca tattgagcca gtagcgcggt gtattatacc
1261 agaccttatt ggtatgggca aatcaggcaa atctggtaat ggttcttata ggttacttga
1321 tcattacaaa tatcttactg catggtttga acttcttaat ttaccaaaga agatcatttt
1381 tgtcggccat gattggggtg cttgtttggc atttcattat agctatgagc atcaagataa
1441 gatcaaagca atagttcacg ctgaaagtgt agtagatgtg attgaatcat gggatgaatg
1501 gcctgatatt gaagaagata ttgcgttgat caaatctgaa gaaggagaaa aaatggtttt
1561 ggagaataac ttcttcgtgg aaaccatgtt gccatcaaaa atcatgagaa agttagaacc
1621 agaagaattt gcagcatatc ttgaaccatt caaagagaaa ggtgaagttc gtcgtccaac
1681 attatcatgg cctcgtgaaa tcccgttagt aaaaggtggt aaacctgacg ttgtacaaat
1741 tgttaggaat tataatgctt atctacgtgc aagtgatgat ttaccaaaaa tgtttattga
1801 atcggaccca ggattctttt ccaatgctat tgttgaaggt gccaagaagt ttcctaatac
1861 tgaatttgtc aaagtaaaag gtcttcattt ttcgcaagaa gatgcacctg atgaaatggg
1921 aaaatatatc aaatcgttcg ttgagcgagt tctcaaaaat gaacaataat tctagagcgg
1981 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
2041 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
2101 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
2161 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
2221 aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
2281 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
2341 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
2401 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
2461 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
2521 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
2581 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
2641 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
2701 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
2761 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
2821 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
2881 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
2941 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
3001 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
3061 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
3121 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
3181 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
3241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
3301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
3361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
3421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
3481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
3541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
3601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
3661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
3721 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
3781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
3841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
3901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
3961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
4021 tggccttttg ctcacatggc tcgac
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后,请先根据产品管壁标签来判断产品形式,并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉(常温运输,存于-20度,90天保质期,请务必转化挑单克隆培养,不要直接使用和测序)
①收到质粒干粉后请先5000rpm离心1min,再加入20μl ddH2O去离子水溶解质粒;
②取1支100μl 感受态于冰上解冻10min,加入2μl质粒,再冰浴30min后,42℃热激60s,不要搅动,再冰浴2min;(从第二步开始均要在超净工作台中无菌操作)
③加入900μl无抗的LB液体培养基,180rpm震荡37℃培养45min;
④6000rpm离心5min,仅留100μl上清液重悬细菌沉淀,并涂布至目标质粒抗性的LB平板上;(可使用本平台的平板涂布专用玻璃珠进行涂布,可以比传统涂布方法获得更多转化子)
⑤将平板正向培养1h,再倒置37℃培养14h。如果要求是30度则培养20h;
(菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态,要先转克隆感受态,重提质粒后再导入表达感受态)
⑥挑取单菌落至LB液体培养基中,加入对应抗生素,220rpm震荡培养14h,根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种(冰袋运输,存于-80℃,保质期90天,请务必划线挑单克隆培养)
四区划线后挑单菌落培养,酵母菌需要先液体复苏再四区划线,再挑单菌落液体培养。
3、穿刺菌种(冰袋运输,存于4℃,保质期7天)
穿刺接种,液体培养后四区划线,再挑单菌落液体培养。
4、菌落平板(冰袋运输,存于4℃,保质期7天)
直接挑取单菌落至液体培养基中。
5、液体质粒(冰袋运输,存于-20℃,保质期90天)
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒(常温运输,存于-20度,90天保质期,请务必转化挑单克隆培养,不要直接使用和测序)
收到货后将滤纸画圈部分剪下放入EP管中,加100ul无菌水将滤纸浸湿并浸泡5min,吸取5ul质粒转化,离心全涂。
二、转化图片
| P4613/pCMV-SPORT6-TPP1(1点突变)人源基因质粒 |
| P4614/pCMV-SPORT6-PANX1-A15C人源基因质粒 |
| P4615/pCMV-SPORT6-ACOX3人源基因质粒 |
| P4616/pCMV-SPORT6-SOCS6人源基因质粒 |
| P4617/pCMV-SPORT6-HLA-DOA人源基因质粒 |
| P4618/pCMV-SPORT6-ADORA2A人源基因质粒 |
| P4619/pCMV-SPORT6-ZBTB22(1点突变)人源基因质粒 |
| P4620/pCMV-SPORT6-ARL8A人源基因质粒 |
| P4621/pCMV-SPORT6-MAGEB1人源基因质粒 |
| P4622/pCMV-SPORT6-MUDENG人源基因质粒 |
| P4623/pCMV-SPORT6-CLIP3人源基因质粒 |
| P4624/pCMV-SPORT6-MRPL32(1同义突变)人源基因质粒 |
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pRL-TK哺乳报告质粒
¥100 - 1000










