Product Information

Transformation of plasmid DNA to competent E. Coli cells

1. Thaw competent cells on ice. 20–200µL per tube
2. Add max. 1-3µL of plasmid
3. Mix very gently!
4. Incubate the tubes on ice for 30 min
5. Heat shock the cells for 45 sec to 2 min at 42°C
6. Place the tubes immediately on ice for at least 2 min
7. Add 800µL of SOC medium to each tube
8. Incubate for 1 hour at 37°C and shake vigorously
9. Spin down briefly and remove most supernatants
10. Resuspend cell pellet with the rest SOC medium in the tube by pipetting
11. Plate out the suspension on a LB agar plate containing the appropriate antibiotic. Incubate the plates overnight at 37°C

LOCUS pBluescript II SK(-) 2959 bp DNA CIRCULAR SYN 19-MAR-2009

DEFINITION

ACCESSION pBluescript II SK(-)

KEYWORDS .

SOURCE Unknown.

ORGANISM Unknown

Unclassified.

REFERENCE 1 (bases 1 to 2959)

AUTHORS Self

JOURNAL Unpublished.

COMMENT SECID/File created by SciEd Central, Scientific & Educational Software

FEATURES Location/Qualifiers

CDS 21..327

/gene="f1(-)origin"

misc_feature 600..643

/gene="T7/M13F"

CDS complement (461..615)

/gene="lacZ"

misc_feature complement (770..826)

/gene="T3/M13R"

misc_feature 1156..1823

/gene="pUC origin"

CDS complement (1974..2831)

/gene="Amp"

BASE COUNT 707 a 755 c 731 g 766 t

ORIGIN

1 CTGACGCGCC CTGTAGCGGC GCATTAAGCG CGGCGGGTGT GGTGGTTACG CGCAGCGTGA

61 CCGCTACACT TGCCAGCGCC CTAGCGCCCG CTCCTTTCGC TTTCTTCCCT TCCTTTCTCG

121 CCACGTTCGC CGGCTTTCCC CGTCAAGCTC TAAATCGGGG GCTCCCTTTA GGGTTCCGAT

181 TTAGTGCTTT ACGGCACCTC GACCCCAAAA AACTTGATTA GGGTGATGGT TCACGTAGTG

241 GGCCATCGCC CTGATAGACG GTTTTTCGCC CTTTGACGTT GGAGTCCACG TTCTTTAATA

301 GTGGACTCTT GTTCCAAACT GGAACAACAC TCAACCCTAT CTCGGTCTAT TCTTTTGATT

361 TATAAGGGAT TTTGCCGATT TCGGCCTATT GGTTAAAAAA TGAGCTGATT TAACAAAAAT

421 TTAACGCGAA TTTTAACAAA ATATTAACGC TTACAATTTG CCATTCGCCA TTCAGGCTGC

481 GCAACTGTTG GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT ATTACGCCAG CTGGCGAAAG

541 GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG GTTTTCCCAG TCACGACGTT

601 GTAAAACGAC GGCCAGTGAA TTGTAATACG ACTCACTATA GGGCGAATTG GGTACCGGGC