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Putra Taufik 11-13-2010 02:19 AM

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.

zargar.shabir 11-16-2010 08:50 PM

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

Putra Taufik 11-17-2010 07:36 AM

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.

admin 11-17-2010 09:23 AM

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.

SebaQ 11-17-2010 05:02 PM

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 =)

Putra Taufik 11-18-2010 09:06 AM

Re: Vector for large insert cloning
 
ok . than you very much to all of you for your help.


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