pCR Plasmids are a family of cloning vectors designed for the efficient cloning of PCR products. These plasmids are part of the TOPO® cloning system developed by Invitrogen (now part of Thermo Fisher Scientific). The pCR plasmids are specifically engineered to facilitate the direct insertion of PCR products into the plasmid vector, taking advantage of the single 3'-A overhangs generated by Taq DNA polymerase during PCR amplification.

Content

pCR Plasmids: Technical Insights

  • Structure and Components:
    • Origin of Replication (ori): pCR plasmids contain the high-copy-number pUC origin of replication, which allows for the production of numerous plasmid copies per bacterial cell.
    • Antibiotic Resistance Gene:
      • ampR: Provides resistance to ampicillin, allowing for the selection of bacteria that have successfully taken up the plasmid.
    • TOPO® Cloning Site: The plasmid contains a linearized vector with a single 3'-thymidine (T) overhang, which is complementary to the 3'-adenine (A) overhangs on PCR products generated by Taq DNA polymerase. This allows for the efficient and direct ligation of PCR products.
    • LacZα Gene: The pCR plasmid includes the coding sequence for the α-peptide of β-galactosidase. Insertion of a PCR product into the TOPO® cloning site disrupts the lacZα gene, allowing for blue/white screening of recombinant colonies. Blue colonies indicate non-recombinant plasmids, while white colonies indicate successful cloning of the PCR product.
    • Multiple Cloning Site (MCS): Located within the lacZα gene, the MCS contains several unique restriction enzyme sites for easy insertion and manipulation of the PCR product.
  • Applications:
    • Cloning of PCR Products: The primary use of pCR plasmids is the cloning of PCR-amplified DNA fragments. The TOPO® cloning technology facilitates the rapid and efficient insertion of PCR products directly into the plasmid vector without the need for additional enzymatic steps.
    • Blue/White Screening: The lacZα complementation system allows for easy identification of recombinant colonies. Bacteria with intact lacZα produce blue colonies on X-gal/IPTG plates, while those with disrupted lacZα due to the insertion of a PCR product produce white colonies.
    • Sequencing: Cloned PCR products in pCR plasmids can be easily sequenced to verify their identity and accuracy. The high copy number of the plasmid ensures sufficient DNA yield for sequencing reactions.
    • Subcloning: Inserted PCR fragments can be excised using the restriction sites in the MCS and subcloned into other vectors for further functional studies, expression, or manipulation.
  • Types of pCR Plasmids:
    • pCR™2.1-TOPO®: A commonly used vector in the pCR family, designed for efficient TOPO® cloning of PCR products.
    • pCR™4-TOPO®: Another variant that includes additional features for more flexible cloning and downstream applications.
    • pCR™-Blunt II-TOPO®: Designed for blunt-end cloning of PCR products generated by proofreading DNA polymerases, which do not add 3'-A overhangs.
  • Experimental Considerations:
    • Transformation Efficiency: Efficient transformation of competent E. coli cells with pCR plasmids is essential. High-efficiency competent cells should be used to maximize the uptake of the plasmid.
    • Antibiotic Selection: Proper antibiotic selection with ampicillin ensures that only bacteria containing the plasmid will grow. It is important to maintain selective pressure throughout the experiment.
    • Verification: After cloning, the presence and orientation of the inserted PCR product should be verified using techniques such as restriction enzyme digestion, PCR, and sequencing.
For Price Contact info@affigen.com 0.0 USD
AffiPLASMID® pCRY2PHR-Linker-BAX (human)
CAT# AFG-GNC-13923
Size: 1 Unit
For Price Contact info@affigen.com 0.0 USD
AffiPLASMID® pPCRScript-ZSWIM8
CAT# AFG-NVF-111352
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-ZNF415
CAT# AFG-NVF-111351
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-UBASH3A
CAT# AFG-NVF-111350
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-SSX2B
CAT# AFG-NVF-111349
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-SEC14L3
CAT# AFG-NVF-111348
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-RFPL2
CAT# AFG-NVF-111347
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-RETNLB
CAT# AFG-NVF-111346
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-RANBP10
CAT# AFG-NVF-111345
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-PTTG2
CAT# AFG-NVF-111344
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-PRKAA2
CAT# AFG-NVF-111343
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-PGLYRP3
CAT# AFG-NVF-111342
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-PDE6H
CAT# AFG-NVF-111341
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-PAEP
CAT# AFG-NVF-111340
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-PAAF1
CAT# AFG-NVF-111339
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-NELL1
CAT# AFG-NVF-111338
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-NAT8B
CAT# AFG-NVF-111337
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-MS4A6E
CAT# AFG-NVF-111336
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-MC3R
CAT# AFG-NVF-111335
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-LTB
CAT# AFG-NVF-111334
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-LIM2
CAT# AFG-NVF-111333
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-LEP
CAT# AFG-NVF-111332
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-LAIR2
CAT# AFG-NVF-111331
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-LACRT
CAT# AFG-NVF-111330
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-KLK4
CAT# AFG-NVF-111329
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-KitLG
CAT# AFG-NVF-111328
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-IL20
CAT# AFG-NVF-111327
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-IFNA6
CAT# AFG-NVF-111326
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-HTR2A
CAT# AFG-NVF-111325
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-HRC
CAT# AFG-NVF-111324
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-HBD
CAT# AFG-NVF-111323
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-GAGE2A
CAT# AFG-NVF-111322
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-GAGE12C
CAT# AFG-NVF-111321
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-GAGE1
CAT# AFG-NVF-111320
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-FPR3
CAT# AFG-NVF-111319
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-FABP2
CAT# AFG-NVF-111318
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CYP4F22
CAT# AFG-NVF-111317
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CRYBB3
CAT# AFG-NVF-111316
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CRYBA4
CAT# AFG-NVF-111315
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CRYBA1
CAT# AFG-NVF-111314
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CRLS1
CAT# AFG-NVF-111313
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CNR2
CAT# AFG-NVF-111312
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CLDN16
CAT# AFG-NVF-111311
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CHRNB3
CAT# AFG-NVF-111310
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CER1
CAT# AFG-NVF-111309
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CELA2B
CAT# AFG-NVF-111308
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CD200R1
CAT# AFG-NVF-111307
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CCL18
CAT# AFG-NVF-111306
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CBLN4
CAT# AFG-NVF-111305
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CAV3
CAT# AFG-NVF-111304
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-CA8
CAT# AFG-NVF-111303
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCRScript-C19orf73
CAT# AFG-NVF-111302
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCR-Script Amp SK (+) -ZNF415
CAT# AFG-NVF-111301
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCR-Script Amp SK (+) -PGLYRP3
CAT# AFG-NVF-111300
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCR-Script Amp SK (+) -IFNB1
CAT# AFG-NVF-111299
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pPCR-Script Amp SK (+) -IFNA6
CAT# AFG-NVF-111298
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCR-Script Amp SK (+) -CD200R1
CAT# AFG-NVF-111297
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCR-Script Amp SK (+) -CCL18
CAT# AFG-NVF-111296
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pPCR-Script Amp SK (+) -AZU1
CAT# AFG-NVF-111295
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pOTB7-NTPCR
CAT# AFG-NVF-109734
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET22B, SA, PcrA
CAT# AFG-NVF-096244
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET22B, PcrA, 2Bchimera
CAT# AFG-NVF-096243
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCRY2PHR-mCherryN1-Linker-BAX
CAT# AFG-NVF-089236
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCRY2PHR-mCherryN1
CAT# AFG-NVF-089235
Size: 2 μg
198.10 198.1 USD
AffiPLASMID® pCRY2PHR-Linker-BAX Plasmid
CAT# AFG-NVF-089234
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-ZNF236
CAT# AFG-NVF-089233
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-ZNF229
CAT# AFG-NVF-089232
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-ZKSCAN2
CAT# AFG-NVF-089231
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-WASHC4
CAT# AFG-NVF-089230
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-USP45
CAT# AFG-NVF-089229
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-USP42
CAT# AFG-NVF-089228
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-TOPBP1 Plasmid
CAT# AFG-NVF-089227
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-TNN
CAT# AFG-NVF-089226
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-TMEM63C
CAT# AFG-NVF-089225
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-TAOK1
CAT# AFG-NVF-089224
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SPHKAP
CAT# AFG-NVF-089223
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SOS2
CAT# AFG-NVF-089222
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SMC1A
CAT# AFG-NVF-089221
Size: 2 μg
561.00 561.0 USD
AffiPLASMID® pCR-XL-TOPO-SLIT1
CAT# AFG-NVF-089220
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SLC6A20
CAT# AFG-NVF-089219
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SLC24A1
CAT# AFG-NVF-089218
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SLC22A3
CAT# AFG-NVF-089217
Size: 2 μg
371.00 371.0 USD
AffiPLASMID® pCR-XL-TOPO-SIPA1L3
CAT# AFG-NVF-089216
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SHROOM2
CAT# AFG-NVF-089215
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SETD2
CAT# AFG-NVF-089214
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SEC31B
CAT# AFG-NVF-089213
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-SEC14L1
CAT# AFG-NVF-089212
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-RPTOR
CAT# AFG-NVF-089211
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-PTPRG
CAT# AFG-NVF-089210
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-PPP1R9A
CAT# AFG-NVF-089209
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-PHRF1
CAT# AFG-NVF-089208
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-OR4F4-4
CAT# AFG-NVF-089207
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-OR4F4
CAT# AFG-NVF-089206
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pCR-XL-TOPO-OR2AT4
CAT# AFG-NVF-089205
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-OR1L8
CAT# AFG-NVF-089204
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-OR10AG1
CAT# AFG-NVF-089203
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-NRAP
CAT# AFG-NVF-089202
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pCR-XL-TOPO-NPC1L1
CAT# AFG-NVF-089201
Size: 2 μg
230.40 230.4 USD
AffiPLASMID® pCR-XL-TOPO-NCOA6
CAT# AFG-NVF-089200
Size: 2 μg
276.00 276.0 USD