pFREE
$113.00
In stock
SKU
PL39988
100μg liquid plasmid(Instantly):$134
| name | pFREE | another name | apFREE |
| Insert Size (bp) | Plasmid host | Escherichia coli | |
| Plasmid uses | Gene editing | Fragment type | CRISPR |
| Fragment species | Prokaryotic resistance | Kan | |
| Selective marker | Fluorescent labeling | ||
| Promoter | Tet | Replicon | ColA |
| Competence | Stbl3 | Growth Temperature | 30°C |
| Vector backbone | 5′ sequencing primer | ||
| 3′ sequencing primer | Copy number | ||
| Induction methodanother name |
1. All products on this platform are only for scientific research and cannot be used for clinical trials;
2. The small package of plasmids is sold at the price of the template, which only guarantees the correct sequencing of the gene part, and does not guarantee the experimental effect such as the expression of the plasmid;
3. Some plasmids have not been completely sequenced even if they are produced by the company, and if the sequence sequencing of key parts is correct, it is also considered to be the correct plasmid;
4. The after-sales period of small package plasmid products is 3 months from the date you receive the product.
5. Plasmid construction is charged at cost, all plasmids will be shared with other experimenters, please inform and pay confidential fees in advance for confidential plasmids;
6. Because scientific research is an activity to explore the unknown, there is great uncertainty, so Biogege does not bear additional joint and several liability under any circumstances;
7. Publication [Chinese papers] please mark: product name (product item number) provided by Biogege;
8. Please mark Product Name (Product Item Number) was obtained from Biogege,China.
Instructions
Plasmid dry powder (stored at minus 20 degrees, shelf life 90 days, transportation at room temperature)
1. After receiving the dry powder plasmid, please centrifuge at 5000rpm for 1min, and then add 20μl of sterile water to dissolve the plasmid;
2. Thaw the corresponding competent on an ice box, take 2 μl of plasmid and add it to 100 μl of competent for 30min;
3. 42 degrees heat shock for 60s, ice bath for 2min, add 900 of antibiotic-free LB medium, and culture at 180rpm shaking for 45min;
4. Centrifuge at 6000rpm for 3min, leave 100μl of supernatant to resuspend, and then coat it on the corresponding resistant LB plate;
5.After inverting overnight, if there are too many colonies, the plasmid will be diluted for transformation, and if there are no colonies, 10ul plasmid will be taken for transformation;
6. Pick single colonies into the corresponding resistant LB liquid medium overnight shaking culture, and extract the plasmid as needed.
Liquid plasmid (minus 20 degrees storage, shelf life 90 days, ice pack transportation).
1. After receiving the liquid plasmid, please shake the preservation tube so that the liquid plasmid does not adsorb on the tube wall;
2.Take an appropriate amount of plasmid according to the experimental needs, and store the rest in a minus 20 degree freezer.
Glycerol strains (stored at minus 80 degrees, shelf life of 90 days, ice pack transportation).
1. After receiving the glycerol strain, please shake the preservation tube so that the bacterial solution does not adsorb on the tube wall;
2. Use the inoculation loop to mark the line in four areas on the solid plate, and add the corresponding resistance if there is resistance;
3. Inverted culture in constant temperature incubator for 12~16h;
4. Be sure to pick a single colony and inoculate it in the liquid medium, and add the corresponding resistance if there is resistance;
5. Shake the shaker and culture overnight, and culture the strains according to the needs of the experiment.
TCCCTATCAG TGATAGAGAT TGACATCCCT ATCAGTGATA GAGATACTGA GCACAGAAGG AGATATACAT GGATAAGAAA TACTGAGCAC TATCAGTGAT AGAGATACTG AGCACAGAAG GAGATATACA TGGATAAGAA ATACTCAATA GGCTTAGATA TCGGCACAAA TAGCGTCGGA TGGGCGGTGA TCACTGATGA ATATAAGGTT CCGTCTAAAA AGTTCAAGGT TCTGGGAAAT ACAGACCGCC ACAGTATCAA AAAAAATCTT ATAGGGGCTC TTTTATTTGA CAGTGGAGAG ACAGCGGAAG CGACTCGTCT CAAACGGACA GCTCGTAGAA GGTATACACG TCGGAAGAAT CGTATTTGTT ATCTACAGGA GATTTTTTCA AATGAGATGG CGAAAGTAGA TGATAGTTTC TTTCATCGAC TTGAAGAGTC TTTTTTGGTG GAAGAAGACA AGAAGCATGA ACGTCATCCT ATTTTTGGAA ATATAGTAGA TGAAGTTGCT TATCATGAGA AATATCCAAC TATCTATCAT CTGCGAAAAA AATTGGTAGA TTCTACTGAT AAAGCGGATT TGCGCTTAAT CTATTTGGCC TTAGCGCATA TGATTAAGTT TCGTGGTCAT TTTTTGATTG AGGGAGATTT AAATCCTGAT AATAGTGATG TGGACAAACT ATTTATCCAG TTGGTACAAA CCTACAATCA ATTATTTGAA GAAAACCCTA TTAACGCAAG TGGAGTAGAT GCTAAAGCGA TTCTTTCTGC ACGATTGAGT AAATCAAGAC GATTAGAAAA TCTCATTGCT CAGCTCCCCG GTGAGAAGAA AAATGGCTTA TTTGGGAATC TCATTGCTTT GTCATTGGGT TTGACCCCTA ATTTTAAATC AAATTTTGAT TTGGCAGAAG ATGCTAAATT ACAGCTTTCA AAAGATACTT ACGATGATGA TTTAGATAAT TTATTGGCGC AAATTGGAGA TCAATATGCT GATTTGTTTT TGGCAGCTAA GAATTTATCA GATGCTATTT TACTTTCAGA TATCCTAAGA GTAAATACTG AAATAACTAA GGCTCCCCTA TCAGCTTCAA TGATTAAACG CTACGATGAA CATCATCAAG ACTTGACTCT TTTAAAAGCT TTAGTTCGAC AACAACTTCC AGAAAAGTAT AAAGAAATCT TTTTTGATCA ATCAAAAAAC GGATATGCAG GTTATATTGA TGGGGGAGCT AGCCAAGAAG AATTTTATAA ATTTATCAAA CCAATTTTAG AAAAAATGGA TGGTACTGAG GAATTATTGG TGAAACTAAA TCGTGAAGAT TTGCTGCGCA AGCAACGGAC CTTTGACAAC GGCTCTATTC CCCATCAAAT TCACTTGGGT GAGCTGCATG CTATTTTGAG AAGACAAGAA GACTTTTATC CATTTTTAAA AGACAATCGT GAGAAGATTG AAAAAATCTT GACTTTTCGA ATTCCTTATT ATGTTGGTCC ATTGGCGCGT GGCAATAGTC GTTTTGCATG GATGACTCGG AAGTCTGAAG AAACAATTAC CCCATGGAAT TTTGAAGAAG TTGTCGATAA AGGTGCTTCA GCTCAATCAT TTATTGAACG CATGACAAAC TTTGATAAAA ATCTTCCAAA TGAAAAAGTA CTACCAAAAC ATAGTTTGCT TTATGAGTAT TTTACGGTTT ATAACGAATT GACAAAGGTC AAATATGTTA CTGAAGGAAT GCGAAAACCA GCATTTCTTT CAGGTGAACA GAAGAAAGCC ATTGTTGATT TACTCTTCAA AACAAATCGA AAAGTAACCG TTAAGCAATT AAAAGAAGAT TATTTCAAAA AAATAGAATG TTTTGATAGT GTTGAAATTT CAGGAGTTGA AGATAGATTT AATGCTTCAT TAGGTACCTA CCATGATTTG CTAAAAATTA TTAAAGATAA AGATTTTTTG GATAATGAAG AAAATGAAGA TATCTTAGAG GATATTGTTT TAACATTGAC CTTATTTGAA GATAGGGAGA TGATTGAGGA AAGACTTAAA ACATATGCTC ACCTCTTTGA TGATAAGGTG ATGAAACAGC TTAAACGTCG CCGTTATACT GGTTGGGGAC GTTTGTCTCG AAAATTGATT AATGGTATTA GGGATAAGCA ATCTGGCAAA ACAATATTAG ATTTTTTGAA ATCAGATGGT TTTGCCAATC GCAATTTTAT GCAGCTGATC CATGATGATA GTTTGACATT TAAAGAAGAC ATTCAAAAAG CACAAGTGTC TGGACAAGGC GATAGTTTAC ATGAACATAT TGCAAATTTA GCTGGTAGCC CTGCTATTAA AAAAGGTATT TTACAGACTG TAAAAGTTGT TGATGAATTG GTCAAAGTAA TGGGGCGGCA TAAGCCAGAA AATATCGTTA TTGAAATGGC ACGTGAAAAT CAGACAACTC AAAAGGGCCA GAAAAATTCG CGAGAGCGTA TGAAACGAAT CGAAGAAGGT ATCAAAGAAT TAGGAAGTCA GATTCTTAAA GAGCATCCTG TTGAAAATAC TCAATTGCAA AATGAAAAGC TCTATCTCTA TTATCTCCAA AATGGAAGAG ACATGTATGT GGACCAAGAA TTAGATATTA ATCGTTTAAG TGATTATGAT GTCGATCACA TTGTTCCACA AAGTTTCCTT AAAGACGATT CAATAGACAA TAAGGTCTTA ACGCGTTCTG ATAAAAATCG TGGTAAATCG GATAACGTTC CAAGTGAAGA AGTAGTCAAA AAGATGAAAA ACTATTGGAG ACAACTTCTA AACGCCAAGT TAATCACTCA ACGTAAGTTT GATAATTTAA CGAAAGCTGA ACGTGGAGGT TTGAGTGAAC TTGATAAAGC TGGTTTTATC AAACGCCAAT TGGTTGAAAC TCGCCAAATC ACTAAGCATG TGGCACAAAT TTTGGATAGT CGCATGAATA CTAAATACGA TGAAAATGAT AAACTTATTC GAGAGGTTAA AGTGATTACC TTAAAATCTA AATTAGTTTC TGACTTCCGA AAAGATTTCC AATTCTATAA AGTACGTGAG ATTAACAATT ACCATCATGC CCATGATGCG TATCTAAATG CCGTCGTTGG AACTGCTTTG ATTAAGAAAT ATCCAAAACT TGAATCGGAG TTTGTCTATG GTGATTATAA AGTTTATGAT GTTCGTAAAA TGATTGCTAA GTCTGAGCAA GAAATAGGCA AAGCAACCGC AAAATATTTC TTTTACTCTA ATATCATGAA CTTCTTCAAA ACAGAAATTA CACTTGCAAA TGGAGAGATT CGCAAACGCC CTCTAATCGA AACTAATGGG GAAACTGGAG AAATTGTCTG GGATAAAGGG CGAGATTTTG CCACAGTGCG CAAAGTATTG TCCATGCCCC AAGTCAATAT TGTCAAGAAA ACAGAAGTAC AGACAGGCGG ATTCTCCAAG GAGTCAATTT TACCAAAAAG AAATTCGGAC AAGCTTATTG CTCGTAAAAA AGACTGGGAT CCAAAAAAAT ATGGTGGTTT TGATAGTCCA ACGGTAGCTT ATTCAGTCCT AGTGGTTGCT AAGGTGGAAA AAGGGAAATC GAAGAAGTTA AAATCCGTTA AAGAGTTACT AGGGATCACA ATTATGGAAA GAAGTTCCTT TGAAAAAAAT CCGATTGACT TTTTAGAAGC TAAAGGATAT AAGGAAGTTA AAAAAGACTT AATCATTAAA CTACCTAAAT ATAGTCTTTT TGAGTTAGAA AACGGTCGTA AACGGATGCT GGCTAGTGCC GGAGAATTAC AAAAAGGAAA TGAGCTGGCT CTGCCAAGCA AATATGTGAA TTTTTTATAT TTAGCTAGTC ATTATGAAAA GTTGAAGGGT AGTCCAGAAG ATAACGAACA AAAACAATTG TTTGTGGAGC AGCATAAGCA TTATTTAGAT GAGATTATTG AGCAAATCAG TGAATTTTCT AAGCGTGTTA TTTTAGCAGA TGCCAATTTA GATAAAGTTC TTAGTGCATA TAACAAACAT AGAGACAAAC CAATACGTGA ACAAGCAGAA AATATTATTC ATTTATTTAC GTTGACGAAT CTTGGAGCTC CCGCTGCTTT TAAATATTTT GATACAACAA TTGATCGTAA ACGATATACG TCTACAAAAG AAGTTTTAGA TGCCACTCTT ATCCATCAAT CCATCACTGG TCTTTATGAA ACACGCATTG ATTTGAGTCA GCTAGGAGGT GACTGAGATC CCATGTTACC GGTATCCTAG GCTGCTGCCA CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT TGCTGAAACC TCAGGCATTT GAGAAGCACA CGGTCACACT GCTTCCGGTA GTCAATAAAC CGGTATTGAC GCCAACGGGT CACACGGGTA CAGTGCTAGC AGAAACATAT CCATGAAATC CCGCCCCGAA TTCATATGTC TAGATTAGAT AAAAGTAAAG TGATTAACAG CGCATTAGAG CTGCTTAATG AGGTCGGAAT CGAAGGTTTA ACAACCCGTA AACTCGCCCA GAAGCTAGGT GTAGAGCAGC CTACATTGTA TTGGCATGTA AAAAATAAGC GGGCTTTGCT CGACGCCTTA GCCATTGAGA TGTTAGATAG GCACCATACT CACTTTTGCC CTTTAGAAGG GGAAAGCTGG CAAGATTTTT TACGTAATAA CGCTAAAAGT TTTAGATGTG CTTTACTAAG TCATCGCGAT GGAGCAAAAG TACATTTAGG TACACGGCCT ACAGAAAAAC AGTATGAAAC TCTCGAAAAT CAATTAGCCT TTTTATGCCA ACAAGGTTTT TCACTAGAGA ATGCATTATA TGCACTCAGC GCTGTGGGGC ATTTTACTTT AGGTTGCGTA TTGGAAGATC AAGAGCATCA AGTCGCTAAA GAAGAAAGGG AAACACCTAC TACTGATAGT ATGCCGCCAT TATTACGACA AGCTATCGAA TTATTTGATC ACCAAGGTGC AGAGCCAGCC TTCTTATTCG GCCTTGAATT GATCATATGC GGATTAGAAA AACAACTTAA ATGTGAAAGT GGGTCTTAAG CAATAGACAT AAGCGGCTAT TTAACGACCC TGCCCTGAAC CGACGACAAG CTGACGACCG GGTCTCCGCA AGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT CTAAATACAT TCAAATATGT ATCCGCTCAT GAATTAATTC TTAGAAAAAC TCATCGAGCA TCAAATGAAA CTGCAATTTA TTCATATCAG GATTATCAAT ACCATATTTT TGAAAAAGCC GTTTCTGTAA TGAAGGAGAA AACTCACCGA GGCAGTTCCA TAGGATGGCA AGATCCTGGT ATCGGTCTGC GATTCCGACT CGTCCAACAT CAATACAACC TATTAATTTC CCCTCGTCAA AAATAAGGTT ATCAAGTGAG AAATCACCAT GAGTGACGAC TGAATCCGGT GAGAATGGCA AAAGTTTATG CATTTCTTTC CAGACTTGTT CAACAGGCCA GCCATTACGC TCGTCATCAA AATCACTCGC ATCAACCAAA CCGTTATTCA TTCGTGATTG CGCCTGAGCG AGACGAAATA CGCGGTCGCT GTTAAAAGGA CAATTACAAA CAGGAATCGA ATGCAACCGG CGCAGGAACA CTGCCAGCGC ATCAACAATA TTTTCACCTG AATCAGGATA TTCTTCTAAT ACCTGGAATG CTGTTTTCCC GGGGATCGCA GTGGTGAGTA ACCATGCATC ATCAGGAGTA CGGATAAAAT GCTTGATGGT CGGAAGAGGC ATAAATTCCG TCAGCCAGTT TAGTCTGACC ATCTCATCTG TAACATCATT GGCAACGCTA CCTTTGCCAT GTTTCAGAAA CAACTCTGGC GCATCGGGCT TCCCATACAA TCGATAGATT GTCGCACCTG ATTGCCCGAC ATTATCGCGA GCCCATTTAT ACCCATATAA ATCAGCATCC ATGTTGGAAT TTAATCGCGG CCTAGAGCAA GACGTTTCCC GTTGAATATG GCTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA TAAACAAATA GGCATGCTAG CGCAGAAACG TCCTAGAAGA TGCCAGGAGG ATACTTAGCA GAGAGACAAT AAGGCCGGAG CGAAGCCGTT TTTCCATAGG CTCCGCCCCC CTGACGAACA TCACGAAATC TGACGCTCAA ATCAGTGGTG GCGAAACCCG ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGATGGCT CCCTCTTGCG CTCTCCTGTT CCCGTCCTGC GGCGTCCGTG TTGTGGTGGA GGCTTTACCC AAATCACCAC GTCCCGTTCC GTGTAGACAG TTCGCTCCAA GCTGGGCTGT GTGCAAGAAC CCCCCGTTCA GCCCGACTGC TGCGCCTTAT CCGGTAACTA TCATCTTGAG TCCAACCCGG AAAGACACGA CAAAACGCCA CTGGCAGCAG CCATTGGTAA CTGAGAATTA GTGGATTTAG ATATCGAGAG TCTTGAAGTG GTGGCCTAAC AGAGGCTACA CTGAAAGGAC AGTATTTGGT ATCTGCGCTC CACTAAAGCC AGTTACCAGG TTAAGCAGTT CCCCAACTGA CTTAACCTTC GATCAAACCG CCTCCCCAGG CGGTTTTTTC GTTTACAGAG CAGGAGATTA CGACGATCGT AAAAGGATCT CAAGAAGATC CTTTACGGAT TCCCGACACC ATCACTCTAG ATTTCAGTGC AATTTATCTC TTCAAATGTA GCACCTGAAG TCAGCCCCAT ACGATATAAG TTGTAATTCT CATGTTAGTC ATGCCCCGCG CCCACCGGAA GGAGCTGACT GGGTTGAAGG CTCTCAAGGG CATCGGTCGA GATCCCGGTG CCTAATGAGT GAGCTAACTT ACATTAATTG CGTTGCGCAC ACCGACTAGC GAAAAAACCC CGCCGAAGCG GGGTTTTTTG CGAAAAAAAG CACCGACTCG GTGCCACTTT TTCAAGTTGA TAACGGACTA GCCTTATTTT AACTTGCTAT GCTGTTTTGA ATGGTTCCGC TAGCACTGTA CCTAGGACTG AGCTAGCCGT CAAAAGCAGC GTATATACCA TGGCCACAAT TCAGCAAATT GTGAACATCA TCACGTTCAT CTTTCCCTGG TTGCCAATGG CCCATTTTCC TGTCAGTAAC GAGAAGGTCG CGAATTCAGG CGCTTTTTAG ACTGGTCGTG TTTTAGAGCT ATGCTGTTTT GAATGGTCCC AAAACATGAA CTAGCGATTA GTCGCTATGA CTTAAGTTTT AGAGCTATGC TGTTTTGAAT GGTCCCAAAA CAACCACACT AGAGAACATA CTGGCTAAAT AGTTTTAGAG CTATGCTGTT TTGAATGGTC CCAAAACGGT TGGACTCAAG ACGATAGTTA CCGGATAGTT TTAGAGCTAT GCTGTTTTGA ATGGTCCCAA AACGGCGAAA CCCGACAGGA CTATAAAGAT ACCGTTTTAG AGCTATGCTG TTTTGAATGG TCCCAAAACA AAAAAAAACC CCGCCCCTGA CAGGGCGGGG TTTTTTTTTT AATTAAAGTC TGGTCCTCGA GTCTGGTTAT AA
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