pCold II
$99.00
In stock
SKU
PL83301
pColdII
100μg liquid plasmid(Instantly):$120
| name | pCold II | another name | apCold II |
| Insert Size (bp) | Plasmid host | Escherichia coli | |
| Plasmid uses | Protein expression | Fragment type | ORF |
| Fragment species | Empty vector | Prokaryotic resistance | Amp |
| Selective marker | Fluorescent labeling | ||
| Promoter | CSPA | Replicon | pBR322 |
| Competence | DH5a | Growth Temperature | 37°C |
| Vector backbone | 5′ sequencing primer | pCold-F：ACGCCATATCGCCGAAAGG | |
| 3′ sequencing primer | pCold-R：GGCAGGGATCTTAGATTCTG | Copy number | high |
| 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.
AAGGAATGGT GTGGCCGATT AATCATAAAT ATGAAAAATA ATTGTTGCAT CACCCGCCAA TGCGTGGCTT AATGCACATC AAATTGTGAG CGGATAACAA TTTGATGTGC TAGCGCATAT CCAGTGTAGT AAGGCAAGTC CCTTCAAGAG TTATCGTTGA TACCCCTCGT AGTGCACATT CCTTTAACGC TTCAAAATCT GTAAAGCACG CCATATCGCC GAAAGGCACA CTTAATTATT AAGAGGTAAT ACACCATGAA TCACAAAGTG CATCATCATC ATCATCATAT GGAGCTCGGT ACCCTCGAGG GATCCGAATT CAAGCTTGTC GACCTGCAGT CTAGATAGGT AATCTCTGCT TAAAAGCACA GAATCTAAGA TCCCTGCCAT TTGGCGGGGA TTTTTTTATT TGTTTTCAGG AAATAAATAA TCGATCGCGT AATAAAATCT ATTATTATTT TTGTGAAGAA TAAATTTGGG TGCAATGAGA ATGCGCAGGC CCTTTCGTCT CGCGCGTTTC GGTGATGACG GTGAAAACCT CTGACACATG CAGCTCCCGG AGACGGTCAC AGCTTGTCTG TAAGCGGATG CCGGGAGCAG ACAAGCCCGT CAGGGCGCGT CAGCGGGTGT TGGCGGGTGT CGGGGCTGGC TTAACTATGC GGCATCAGAG CAGATTGTAC TGAGAGTGCA CCATAAAATT GTAAACGTTA ATATTTTGTT AAAATTCGCG TTAAATTTTT GTTAAATCAG CTCATTTTTT AACCAATAGG CCGAAATCGG CAAAATCCCT TATAAATCAA AAGAATAGCC CGAGATAGGG TTGAGTGTTG TTCCAGTTTG GAACAAGAGT CCACTATTAA AGAACGTGGA CTCCAACGTC AAAGGGCGAA AAACCGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC ACCCAAATCA AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA ACCCTAAAGG GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA AGGAAGGGAA GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA GCGGTCACGC TGCGCGTAAC CACCACACCC GCCGCGCTTA ATGCGCCGCT ACAGGGCGCG TACTATGGTT GCTTTGACGT ATGCGGTGTG AAATACCGCA CAGATGCGTA AGGAGAAAAT ACCGCATCAG GCGTCAGGTG GCACTTTTCG GGGAAATGTG CGCGGAACCC CTATTTGTTT ATTTTTCTAA ATACATTCAA ATATGTATCC GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT TGAAAAAGGA AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG GCATTTTGCC TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA AGATGCTGAA GATCAGTTGG GTGCACGAGT GGGTTACATC GAACTGGATC TCAACAGCGG TAAGATCCTT GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA ATGATGAGCA CTTTTAAAGT TCTGCTATGT GGCGCGGTAT TATCCCGTAT TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT TCTCAGAATG ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG ACAGTAAGAG AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC GGCCAACTTA CTTCTGACAA CGATCGGAGG ACCGAAGGAG CTAACCGCTT TTTTGCACAA CATGGGGGAT CATGTAACTC GCCTTGATCG TTGGGAACCG GAGCTGAATG AAGCCATACC AAACGACGAG CGTGACACCA CGATGCCTGT AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA CTACTTACTC TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA GGACCACTTC TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA ATCTGGAGCC GGTGAGCGTG GGTCTCGCGG TATCATTGCA GCACTGGGGC CAGATGGTAA GCCCTCCCGT ATCGTAGTTA TCTACACGAC GGGGAGTCAG GCAACTATGG ATGAACGAAA TAGACAGATC GCTGAGATAG GTGCCTCACT GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT ATACTTTAGA TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTTCTTCTA GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATAGTCAT GCCCCGCGCC CACCGGAAGG AGCTGACTGG GTTGAAGGCT CTCAAGGGCA TCGGTCGAGA TCCCGGTGCC TAATGAGTGA GCTAACTTAC ATTAATTGCG TTGCGCTCAC TGCCCGCTTT CCAGTCGGGA AACCTGTCGT GCCAGCTGCA TTAATGAATC GGCCAACGCG CGGGGAGAGG CGGTTTGCGT ATTGGGCGCC AGGGTGGTTT TTCTTTTCAC CAGTGAGACG GGCAACAGCT GATTGCCCTT CACCGCCTGG CCCTGAGAGA GTTGCAGCAA GCGGTCCACG CTGGTTTGCC CCAGCAGGCG AAAATCCTGT TTGATGGTGG TTAACGGCGG GATATAACAT GAGCTGTCTT CGGTATCGTC GTATCCCACT ACCGAGATAT CCGCACCAAC GCGCAGCCCG GACTCGGTAA TGGCGCGCAT TGCGCCCAGC GCCATCTGAT CGTTGGCAAC CAGCATCGCA GTGGGAACGA TGCCCTCATT CAGCATTTGC ATGGTTTGTT GAAAACCGGA CATGGCACTC CAGTCGCCTT CCCGTTCCGC TATCGGCTGA ATTTGATTGC GAGTGAGATA TTTATGCCAG CCAGCCAGAC GCAGACGCGC CGAGACAGAA CTTAATGGGC CCGCTAACAG CGCGATTTGC TGGTGACCCA ATGCGACCAG ATGCTCCACG CCCAGTCGCG TACCGTCTTC ATGGGAGAAA ATAATACTGT TGATGGGTGT CTGGTCAGAG ACATCAAGAA ATAACGCCGG AACATTAGTG CAGGCAGCTT CCACAGCAAT GGCATCCTGG TCATCCAGCG GATAGTTAAT GATCAGCCCA CTGACGCGTT GCGCGAGAAG ATTGTGCACC GCCGCTTTAC AGGCTTCGAC GCCGCTTCGT TCTACCATCG ACACCACCAC GCTGGCACCC AGTTGATCGG CGCGAGATTT AATCGCCGCG ACAATTTGCG ACGGCGCGTG CAGGGCCAGA CTGGAGGTGG CAACGCCAAT CAGCAACGAC TGTTTGCCCG CCAGTTGTTG TGCCACGCGG TTGGGAATGT AATTCAGCTC CGCCATCGCC GCTTCCACTT TTTCCCGCGT TTTCGCAGAA ACGTGGCTGG CCTGGTTCAC CACGCGGGAA ACGGTCTGAT AAGAGACACC GGCATACTCT GCGACATCGT ATAACGTTAC TGGTTTCACA TTCACCACCC TGAATTGACT CTCTTCCGGG CGCTATCATG CCATACCGCG AAAGGTTTTG CGCCATTCGA TGGTGTCCGG GATCTCGACG CTCTCCCTTA TGCGACTCCT GCATTAGGAA GCAGCCCAGT AGTAGGTTGA GGCCGTTGAG CACCGCCGCC GC
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