Lab Notes: Recombinant DNA Technology Archives - Laboratory Notes https://www.laboratorynotes.com/category/lab-notes-recombinant-dna-technology/ Mon, 05 May 2025 13:36:42 +0000 en hourly 1 https://wordpress.org/?v=6.9 Factors Influencing Plasmid Host-Range Specificity in Bacteria https://www.laboratorynotes.com/factors-influencing-plasmid-host-range-specificity-in-bacteria/ https://www.laboratorynotes.com/factors-influencing-plasmid-host-range-specificity-in-bacteria/#respond Fri, 18 Apr 2025 13:20:41 +0000 https://www.laboratorynotes.com/?p=17880 The host-range specificity of a plasmid refers to the spectrum of bacterial species in which the plasmid can successfully replicate...

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]]> https://www.laboratorynotes.com/factors-influencing-plasmid-host-range-specificity-in-bacteria/feed/ 0 Plasmid DNA Vs Chromosomal DNA https://www.laboratorynotes.com/plasmid-dna-vs-chromosomal-dna/ https://www.laboratorynotes.com/plasmid-dna-vs-chromosomal-dna/#respond Sun, 13 Apr 2025 01:46:18 +0000 https://www.laboratorynotes.com/?p=17727 Criteria Plasmid DNA Chromosomal DNA Remarks Definition Small, circular, double-stranded extrachromosomal DNA molecule found mainly in bacteria and some eukaryotes...

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]]> https://www.laboratorynotes.com/plasmid-dna-vs-chromosomal-dna/feed/ 0 Semi-Quantitative Polymerase Chain Reaction (sqPCR) https://www.laboratorynotes.com/semi-quantitative-polymerase-chain-reaction-sqpcr/ https://www.laboratorynotes.com/semi-quantitative-polymerase-chain-reaction-sqpcr/#respond Tue, 25 Mar 2025 12:09:50 +0000 https://www.laboratorynotes.com/?p=17096 Semi-Quantitative Polymerase Chain Reaction (sqPCR) is a conventional PCR-based technique used to estimate the relative abundance of nucleic acids in...

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]]> https://www.laboratorynotes.com/semi-quantitative-polymerase-chain-reaction-sqpcr/feed/ 0 SYBR Green-based Quantitative PCR (qPCR) https://www.laboratorynotes.com/sybr-green-based-quantitative-pcr-qpcr/ https://www.laboratorynotes.com/sybr-green-based-quantitative-pcr-qpcr/#respond Tue, 25 Mar 2025 11:32:47 +0000 https://www.laboratorynotes.com/?p=17085 SYBR Green-based Quantitative PCR (qPCR) is a widely utilized method for the detection and quantification of nucleic acids in real-time...

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]]> https://www.laboratorynotes.com/sybr-green-based-quantitative-pcr-qpcr/feed/ 0 Sequence-specific Probe-based Quantitative PCR (qPCR) https://www.laboratorynotes.com/sequence-specific-probe-based-quantitative-pcr-qpcr/ https://www.laboratorynotes.com/sequence-specific-probe-based-quantitative-pcr-qpcr/#respond Tue, 25 Mar 2025 11:32:41 +0000 https://www.laboratorynotes.com/?p=17088 Sequence-specific probe-based quantitative PCR (qPCR) is an advanced real-time PCR technique that enables highly specific and sensitive quantification of nucleic...

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]]> https://www.laboratorynotes.com/sequence-specific-probe-based-quantitative-pcr-qpcr/feed/ 0 SYBR Green-based qPCR Vs Sequence-specific Probe-based qPCR https://www.laboratorynotes.com/sybr-green-based-qpcr-vs-sequence-specific-probe-based-qpcr/ https://www.laboratorynotes.com/sybr-green-based-qpcr-vs-sequence-specific-probe-based-qpcr/#respond Tue, 25 Mar 2025 11:32:06 +0000 https://www.laboratorynotes.com/?p=17090 Criteria SYBR Green qPCR Sequence-Specific Probe-Based qPCR Remarks Detection Chemistry Non-specific DNA-binding dye (SYBR Green I) binds to any double-stranded...

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]]> https://www.laboratorynotes.com/sybr-green-based-qpcr-vs-sequence-specific-probe-based-qpcr/feed/ 0 Semi-quantitative PCR (sqPCR) Vs Quantitative PCR (qPCR) https://www.laboratorynotes.com/semi-quantitative-pcr-sqpcr-vs-quantitative-pcr-qpcr/ https://www.laboratorynotes.com/semi-quantitative-pcr-sqpcr-vs-quantitative-pcr-qpcr/#respond Tue, 25 Mar 2025 11:31:53 +0000 https://www.laboratorynotes.com/?p=17092 Criteria Semi-Quantitative PCR (sqPCR) Quantitative PCR (qPCR) Remarks Purpose Estimates relative abundance of target nucleic acids by comparing band intensities...

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]]> https://www.laboratorynotes.com/semi-quantitative-pcr-sqpcr-vs-quantitative-pcr-qpcr/feed/ 0 Quantitative Polymerase Chain Reaction (qPCR) https://www.laboratorynotes.com/quantitative-polymerase-chain-reaction-qpcr/ https://www.laboratorynotes.com/quantitative-polymerase-chain-reaction-qpcr/#respond Mon, 24 Mar 2025 13:43:39 +0000 https://www.laboratorynotes.com/?p=17083 Quantitative Polymerase Chain Reaction (qPCR), also known as real-time PCR, is an advanced molecular biology technique designed for the quantification...

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]]> https://www.laboratorynotes.com/quantitative-polymerase-chain-reaction-qpcr/feed/ 0 Preparing Culture of Escherichia coli for Plasmid Isolation https://www.laboratorynotes.com/preparing-culture-of-escherichia-coli-for-plasmid-isolation/ https://www.laboratorynotes.com/preparing-culture-of-escherichia-coli-for-plasmid-isolation/#respond Thu, 29 Jun 2023 07:18:20 +0000 https://www.laboratorynotes.com/?p=13591 Preparing a culture of plasmid-harboring Escherichia coli is the starting point in all plasmid isolation protocols. Suboptimal culture conditions and...

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]]> https://www.laboratorynotes.com/preparing-culture-of-escherichia-coli-for-plasmid-isolation/feed/ 0 Preparation of Culture of Escherichia coli for Plasmid Minipreparation https://www.laboratorynotes.com/preparation-of-culture-of-escherichia-coli-for-plasmid-minipreparation/ https://www.laboratorynotes.com/preparation-of-culture-of-escherichia-coli-for-plasmid-minipreparation/#respond Sun, 02 Apr 2023 15:34:39 +0000 https://www.laboratorynotes.com/?p=13368 A small-scale plasmid isolation procedure, the miniprep, yields a sufficient amount of plasmid for the screening of clones by restriction...

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]]> https://www.laboratorynotes.com/preparation-of-culture-of-escherichia-coli-for-plasmid-minipreparation/feed/ 0 Restriction enzyme-free Cloning https://www.laboratorynotes.com/restriction-enzyme-free-cloning/ https://www.laboratorynotes.com/restriction-enzyme-free-cloning/#comments Thu, 04 Nov 2021 13:26:30 +0000 https://www.laboratorynotes.com/?p=6763 In contrast to conventional cloning strategies, Restriction enzyme-free cloning (also called Restriction-free cloning) involves no treatment of DNA fragments to...

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]]> https://www.laboratorynotes.com/restriction-enzyme-free-cloning/feed/ 1 T4 DNA Ligase https://www.laboratorynotes.com/t4-dna-ligase/ https://www.laboratorynotes.com/t4-dna-ligase/#comments Tue, 02 Nov 2021 14:38:26 +0000 https://www.laboratorynotes.com/?p=6799 T4 DNA ligase is a highly efficient enzyme derived from the T4 bacteriophage that catalyzes the formation of phosphodiester bonds...

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]]> https://www.laboratorynotes.com/t4-dna-ligase/feed/ 1 TA Cloning https://www.laboratorynotes.com/ta-cloning/ https://www.laboratorynotes.com/ta-cloning/#comments Thu, 07 Oct 2021 04:16:58 +0000 http://www.laboratorynotes.com/?p=4752 TA cloning involves base pairing of adenine (A) and thymine (T)  (complementary base pairs) that result in the joining of...

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]]> https://www.laboratorynotes.com/ta-cloning/feed/ 1 Molecular Cloning (DNA Cloning) https://www.laboratorynotes.com/molecular-cloning-dna-cloning/ https://www.laboratorynotes.com/molecular-cloning-dna-cloning/#respond Thu, 07 Oct 2021 04:16:05 +0000 http://www.laboratorynotes.com/?p=4727 Molecular cloning or DNA cloning refers to a set of methods that involve in generating identical copies of a piece...

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]]> https://www.laboratorynotes.com/molecular-cloning-dna-cloning/feed/ 0 Pfu DNA Polymerase https://www.laboratorynotes.com/pfu-dna-polymerase/ https://www.laboratorynotes.com/pfu-dna-polymerase/#respond Wed, 06 Oct 2021 04:16:01 +0000 http://www.laboratorynotes.com/?p=4735 Pfu DNA polymerase is a high-fidelity, thermostable DNA-dependent DNA polymerase originally isolated from the hyperthermophilic archaeon Pyrococcus furiosus (Lundberg et...

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]]> https://www.laboratorynotes.com/pfu-dna-polymerase/feed/ 0 KOD DNA Polymerase https://www.laboratorynotes.com/kod-dna-polymerase/ https://www.laboratorynotes.com/kod-dna-polymerase/#respond Wed, 06 Oct 2021 04:15:58 +0000 http://www.laboratorynotes.com/?p=4843 Also known as KOD HiFi DNA Polymerase High-fidelity thermostable DNA polymerase for error-free polymerase chain reaction. Originally isolated from Thermococcus...

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]]> https://www.laboratorynotes.com/kod-dna-polymerase/feed/ 0 Phusion DNA Polymerase https://www.laboratorynotes.com/phusion-dna-polymerase/ https://www.laboratorynotes.com/phusion-dna-polymerase/#respond Wed, 06 Oct 2021 04:15:49 +0000 http://www.laboratorynotes.com/?p=4736 Overview: High-fidelity thermostable DNA polymerase for error-free polymerase chain reaction. Possess 3′ to 5′ exonuclease activity for proofreading activity Produces...

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]]> https://www.laboratorynotes.com/phusion-dna-polymerase/feed/ 0 Principle of Alkaline Lysis Method https://www.laboratorynotes.com/principle-of-alkaline-lysis-method/ https://www.laboratorynotes.com/principle-of-alkaline-lysis-method/#comments Mon, 09 Aug 2021 05:46:41 +0000 http://www.laboratorynotes.com/?p=3160 The alkaline lysis method selectively purifies plasmid DNA from other cellular components of the bacterial cells including chromosomal DNA. Controlled...

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]]> https://www.laboratorynotes.com/principle-of-alkaline-lysis-method/feed/ 1 Plasmid Isolation by Alkaline Lysis Method https://www.laboratorynotes.com/plasmid-isolation-by-alkaline-lysis-method/ https://www.laboratorynotes.com/plasmid-isolation-by-alkaline-lysis-method/#comments Sun, 08 Aug 2021 06:08:03 +0000 http://www.laboratorynotes.com/?p=3147 Alkaline lysis method of plasmid isolation was originally developed by Birnboim & Doly (1979). In this procedure, bacteria harbouring the desired...

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]]> https://www.laboratorynotes.com/plasmid-isolation-by-alkaline-lysis-method/feed/ 1 Protocol – Growing Large Volume of Liquid Culture of Escherichia coli for Large-Scale Plasmid Isolation https://www.laboratorynotes.com/protocol-growing-large-volume-of-liquid-culture-of-escherichia-coli-for-large-scale-plasmid-isolation/ https://www.laboratorynotes.com/protocol-growing-large-volume-of-liquid-culture-of-escherichia-coli-for-large-scale-plasmid-isolation/#comments Tue, 03 Aug 2021 04:50:50 +0000 http://www.laboratorynotes.com/?p=2406 Large-scale isolation of plasmid requires a large volume of E. coli culture. Large scale plasmid isolation procedures are termed, midiprep (25 – 50 ml starting culture volume) and maxiprep (100 – 500 ml starting culture volume). A starter culture is initially prepared by inoculating a colony in a small volume (2 – 10 ml) of culture medium. Large culture volume is prepared by diluting starter culture in a ratio of 1: 100 to 1: 1000 in the growth medium.

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