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Vector for large insert cloning

Vector for large insert cloning - Molecular Cloning Forum

Vector for large insert cloning - DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.


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  #1  
Old 11-13-2010, 02:19 AM
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Default Vector for large insert cloning



Good day to everybody, i would like to clone a few PCR products of 4-5kb size, may i know what vector can clone the large insert fragment effectively and efficiently. Or if you have any other suggestions please let me know. Thank you very much.
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Old 11-16-2010, 08:50 PM
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Default Re: Vector for large insert cloning

hi,

I would like to know what gene you want to clone. There is lot number of vectors available for this thing but provide information related protein what the protein is membrane , cytosolic , becterial, or nuclear. and restriction site you want to use

Thank you
shabir
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Old 11-17-2010, 07:36 AM
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Default Re: Vector for large insert cloning

thank you very much for your concern. for your information, my purpose is to clone the sequences and use for sequencing and perservation only. the vector i m looking for is clonning vector not expression vector. Do you know what cloning vector is suitable for my case?

thank you.
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Old 11-17-2010, 09:23 AM
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Default Re: Vector for large insert cloning

I have had no issues cloning with TOPO cloning kits and other sub-cloning vectors even that large.

Just use the ligation calculator located on this site to calculate the ratio of DNA insert to vector for your sizes as this is critical to success.
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Old 11-17-2010, 05:02 PM
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Default Re: Vector for large insert cloning

I've used pGEM-T, pTOPO, pJET and pBLUESCRIPT with no problems for construct as large as those you want to clone =)
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Old 11-18-2010, 09:06 AM
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Default Re: Vector for large insert cloning

ok . than you very much to all of you for your help.
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