In-Fusion SMARTer Directional cDNA Library Construction Kit

2015-01-13

azim58 - In-Fusion SMARTer Directional cDNA Library Construction Kit

In-Fusion® SMARTer™ Directional cDNA Library Construction Kit
Cat # 634933

Manual
http://www.clontech.com/US/Products/cDNA_Synthesis_and_Library_Construction
/cDNA_Library_Construction/ibcGetAttachment.jsp?cItemId=17494&amp;fileId=64
30754&amp;sitex=10020:22372:US

This kit makes use of the pSMART2IFD vector

Sequencing primers for the In-Fusion SMARTer Directional cDNA Library
Construction Kit

In-Fusion Sequences (for some reason the manual displays the plasmid so
that the Lac promoter is on the right side and bottom strand of the
inserted sequence)
left In-Fusion sequence (furthest from Lac promoter)
CGGGGTACGATGAGA
right In-Fusion sequence (closest to Lac promoter)
TTGATACCACTGCTT

2-11-13
This kit is pretty much spent and cannot be used any longer. There is
little to no 3' CDS primer left, only about 1 uL of SMARTer V Oligo, and
only about 1 uL of 5X 1st strand buffer.
There does seem to be quite a lot of 5' PCR Primer IIA though. I think
there could even be as much as 100 uL

SMARTer V Oligo

Plans to modify this vector to add a poly T slip site so that all 3
reading frames of the inserted genes are transcribed and translated can
be found here:
http://www.azim58.wikispaces.net/Designing+primers+to+add+poly+T+8-5-12
Note that as of 12-4-12 we no longer plan to make this modification.

===========================================================================
8-31-12

I (or Mara) will use random hexamer primers to amplify the 1st strand
cDNA.
random hexamer primers for human tumor cDNA library

===========================================================================
Note about reverse transcriptase of In-Fusion SMARTer Directional cDNA
Library Construction Kit 1-14-13

azim58wiki: