What is the protocol for cell free method of cloning?

More Maimuna Qazi's questions See All

Why there is continuous need to genetically engineer a yeast that is already an industrially high-yielding strain??

It is generally observed that scientists and researchers worldwide always try to genetically engineer wild-type hyper-producer (high-yielding) strains that already produce enough/excellent...

31 August 2020 4,104 3 View

Differences in Load controlled and displacement controlled conditions ?

Why is that load controlled and displacement controlled boundary conditions give different results for same meshing and boundary restraints ? I specify time at the end of load step as 1 and no of...

13 February 2019 8,670 6 View

Selection of node from a subset of nodes corresponding to maximum displacement in one direction?

How to select a node from a set of selected nodes and get the node number corresponding to maximum value of say y displacement. It should also take the mod (for selection of maximum negative...

27 January 2019 5,097 1 View

How to SELECT THE keypoints associated with area numbers in APDL?

I have selected a subset of areas by ASEL,S,LOC,Y,0 APLOT With the selected areas I have counted them via *GET,A,AREA,,COUNT then I have defined an array via *DIM,B,ARRAY,1,AREA This creates an...

22 July 2018 4,456 4 View

Solar and generator Hybrid power Generation?

i want to make hybrid power source of solar panel and Diesel Generator can any one help me how it will be done help means via matlab coding examples and suggestions the detail file is attached.

22 February 2016 8,277 3 View

Can anyone recommend a protocol to obtain lysates from cells encapsulated in hydrogels?

I am interested in doing a western blot to detect expression of focal adhesion proteins in cells that are encapsulated in alginate hydrogels. The problem seems to be the infiltration of e.g. SDS...

06 February 2016 1,294 7 View

How can i integrate solar energy and Diesel Generator ?

How can i integrate solar energy and Diesel Generator to make it one source of energy i.e micro grid and then use for my must run or regular load,comments and code segment will be appreciated.

02 February 2016 3,894 4 View

Electroless Ni/ Immersion gold plating solder

Does anyone know of any literature on Electroless Ni Immersion gold soldering using Pb containing solders.

08 August 2013 376 1 View

Literature on high temperature solders

I am looking for literature on high temperature solders, also containing Pb. Any suggestions will be highly appreciated.

08 August 2013 4,282 3 View

Is it possible to get orientation theta values from edge detection using contourlet transform? (like sobel operator).

I am new to contourlet transformation. I have seen the pyramid structure of contourlets, when applied to an image. My problem is getting one matrix to result in edge detection, so I may be able to...

29 January 2013 651 2 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Can we distinguish the type of free fluid inside the pelvic and abdominal cavities whether blood or serous fluid using the ultrasound?

during ultrasound examination of acute abdomen, sometimes we see free fluid , can we know the type of fluid ,whether blood or not , using ultrasound only? if yes , what are the sonographic...

01 March 2021 7,476 4 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Can anyone help me with the administration of the "Mind the Gap" Scale for satisfaction with transition to adult care?

We are working on a study focused on adolescents' transition to adult care and are looking for a validated measure for satisfaction with the transition. So far the only measure that we have came...

28 February 2021 2,338 1 View

How to calculate Ligand-active site distance in a ligand-protein complex from MD simulation?

I have run a MD simulation of a ligand-protein complex in GROMACS using CHARMM forcefield. Reviewer has asked me to calculate Ligand-activesite distance and binding free energy of ligand with...

27 February 2021 4,703 1 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Is there an open source free software for text mining and sentiment analysis?

I am looking for a free software for text mining and sentiment analysis for my research on customer review mining (it involves calculating polarity of attributes,opinion oriented information...

25 February 2021 1,920 6 View

Which are the journals for free paper publication?

A bunch of flower for everybody. I would like to ask about the list of journals which we can publish our paper for free. Thanks in advance. Sincerely Walid

25 February 2021 9,790 3 View

Suggestions for occupational stress surveys?

I am currently writing a masters thesis on reasons for occupational stress in the early childhood classroom, and want to include a short survey assessing participants work-related stress levels. I...

24 February 2021 8,678 2 View

Malcolm Nobre

Hello Maimuna

Please refer to the research article below for more information on cell free method of cloning.

PNAS 2005, 102(48), 17332-17336

https://doi.org/10.1073/pnas.0508809102

Best Wishes.

Lukasz Kuryk

This might be useful:

https://www.oriciro.com/assets/images/product/pdf/OriCiro.Cell-Free.Cloning.System.Manual.OTHER_COUNTRIES.pdf

or Article Cell-free cloning using multiply-primed rolling circle ampli...